Lipopeptides as antibacterial agents

ABSTRACT

The present invention relates to novel lipopeptide compounds. The invention also relates to pharmaceutical compositions of these compounds and methods of using these compounds as antibacterial compounds. The invention also relates to methods of producing these novel lipopeptide compounds and intermediates used in producing these compounds.

FIELD OF THE INVENTION

The present invention relates to novel lipopeptide compounds. Theinvention also relates to pharmaceutical compositions of these compoundsand methods of using these compounds as antibacterial compounds. Theinvention also relates to methods of producing these novel lipopeptidecompounds and intermediates used in producing these compounds.

BACKGROUND OF THE INVENTION

The rapid increase in the incidence of gram-positiveinfections—including those caused by resistant bacteria—has sparkedrenewed interest in the development of novel classes of antibiotics. Aclass of compounds which have shown potential as useful antibioticsincludes the A-21978C lipopeptides described in, for example, U.S. Pat.Nos. RE 32,333; RE 32,455; RE 32,311; RE 32,310; U.S. Pat. Nos.4,482,487; 4,537,717; and 5,912,226. Daptomycin, a member of this class,has potent bactericidal activity in vitro and in vivo against clinicallyrelevant gram-positive bacteria that cause serious and lifethreateningdiseases. These bacteria include resistant pathogens, such asvancomycinresistant enterococci (VRE), methicillin resistantStaphylococcus aureus (MRSA), glycopeptide intermediate susceptibleStaphylococcus aureus (GISA), coagulase-negative staphylococci (CNS),and penicillin-resistant Streptococcus pneumoniae (PRSP), for whichthere are few therapeutic alternatives. See, e.g., Tally et al., 1999,Exp. Opin. Invest Drugs 8:1223-1238.

Despite the promise that antibacterial agents such as daptomycin offer,the need for novel antibiotics continues. Many pathogens have beenrepeatedly exposed to commonly-used antibiotics. This exposure has ledto the selection of variant antibacterial strains resistant to a broadspectrum of antibiotics The loss of potency and effectiveness of anantibiotic caused by resistant mechanisms renders the antibioticineffective and consequently can lead to life-threatening infectionsthat are virtually untreatable. As new antibiotics come to marketpathogens may develop resistance or intermediate resistance to these newdrugs, effectively creating a need for a stream of new antibacterialagents to combat these emerging strains. In addition compounds thatexhibit bacteriacidal activity would offer advantages over presentbacteriastatic compounds. Thus, novel synthetic antibacterial agentswould be expected to be useful to treat not only “natural” pathogens,but also intermediate drug resistant and drug resistant pathogensbecause the pathogen has never been exposed to the novel antibacterialagent. Additionally, new antibacterial agents may exhibit differentialeffectiveness against different types of pathogens.

SUMMARY OF THE INVENTION

The present invention addresses this problem by providing novellipopeptide compounds which have antibacterial activity against a broadspectrum of bacteria, including drug-resistant bacteria. Further, thecompounds of the present invention exhibit bacteriacidal activity.

The present invention comprises, in one aspect, antibacterial compoundsof Formula I:

wherein X and X″ are independently selected from C═O, C═S, C═NH,C═NR^(X), S═O or SO₂;

wherein n is 0 or 1;

wherein R^(X) is selected from alkyl, alkenyl, alkynyl, aryl,heteroaryl, cycloalkyl, heterocyclyl, hydroxyl, alkoxy, carboxy orcarboalkoxy,

wherein B is X″R^(Y), H, alkyl, alkenyl, alkynyl, aryl, heteroaryl,cycloalkyl or heterocyclyl; and

wherein R^(Y) is selected from hydrido, alkyl, alkenyl, alkynyl, aryl,heteroaryl, cycloalkyl, heterocyclyl or hydroxyl.

In one aspect, A is H, NH₂, NHR^(A), NR^(A)R^(B), heteroaryl, cycloalkylor heterocyclyl;

wherein R^(A) and R^(B) are independently selected from alkyl, alkenyl,alkynyl, aryl, heteroaryl, cycloalkyl, heterocyclyl or carboalkoxy;

wherein when n is 0, then A is additionally selected from

wherein each R⁵⁰-R⁵³ is independently selected from (C₁-C₁₅) alkyl,

provided that when B is H and X is C═O, then A is other than

(a) a pyridinyl ring substituted with a single NHC(O)R^(D) substitutentor

(b) a (C₅-C₆) saturated cycloalkyl ring substituted with a singleNHC(O)R^(D) substitutent, wherein R^(D) is (C₁-C₁₇) unsubstituted alkylor (C₂C₁₇) unsubstituted alkenyl; and

when B is H and n is 0, then A is not H.

In another aspect, A is aryl;

provided that when B is H and X is C═O, then A is other than a phenylring substituted with either:

(a) —O—((C₈-C₁₅) unsubstituted alkyl), wherein said phenyl ring may befurther optionally substituted with one substituent selected from halo,nitro, (C₁-C₃) alkyl, hydroxyl, (C₁-C₃) alkoxy or (C₁-C₃) alkylthio; or

(b) —NHC(O)R^(D), wherein the phenyl ring may be further optionallysubstituted with 1-2 substituents independently selected from amino,nitro, (C₁-C₃) alkyl, hydroxyl, (C₁-C₃) alkoxy, halo, mercapto, (C₁-C₃)alkylthio, carbamyl or (C₁-C₃) alkylcarbamyl; wherein R^(D) is asdefined previously.

In a third aspect of the invention, A is alkyl, alkenyl, alkynyl, alkoxyor aryloxy;

provided that when B is H and X is C═O, then A is other than

(a) —(C₁-C₁₆ unsubstituted alkyl)-NH₂;

(b) —(C₁-C₁₀ unsubstituted alkyl)-NHC(O)R^(D), wherein R^(D) is asdefined previously;

(c) —(C₁-C₁₈)-alkyl, optionally substituted with up to one hydroxyl,carboxyl, or C₁-C₃ alkoxy, or one to three halo substituents;

(d) —(C₄-C₁₈)-unsubstituted alkenyl;

wherein R⁵⁴ is selected from C₁-C₁₇unsubstituted alkyl orC₂-C₁₇-unsubstituted alkenyl; wherein R⁵⁵ is selected from hydroxyethyl,hydroxymethyl, mercaptomethyl, mercaptoethyl, methylthioethyl,2-thienyl, 3-indolemethyl, phenyl optionally substituted with a groupselected from halo, nitro, C₁-C₃-unsubstituted alkyl, hydroxy,C₁-C₃-unsubstituted alkoxy, C₁-C₃-unsubsituted alkylthio, carbamyl orC₁-C₃ unsubstituted alkylcarbamyl; or benzyl optionally substituted witha group selected from halo, nitro, C₁-C₃-unsubstituted alkyl, hydroxy,C₁-C₃-unsubstituted alkoxy, C₁-C₃-unsubsituted alkylthio, carbamyl orC₁-C₃ unsubstituted alkylcarbamyl; wherein t is 0 or 1 and wherein u isan integer from 1-3; and

when B is H and X is C═O, then X, together with A, does not form acarbamate amino protecting group; and

when B is H and n is 0, then A is other than C₄C₁₄ unsubstituted alkyl.

In a fourth aspect, B and A together form a 5-7 membered heterocyclic orheteroaryl ring.

Wherein R¹ is

wherein X′ and X′″ are independently selected from C═O, C═S, C═NH,C═NR^(X′), S═O or SO₂;

wherein m is 0 or 1;

wherein R^(X′) is selected from alkyl, alkenyl, alkynyl, aryl,heteroaryl, cycloalkyl, heterocyclyl, hydroxyl, alkoxy, carboxy orcarboalkoxy;

wherein B′ is X′″ R^(Y′), H, alkyl, alkenyl, alkynyl, aryl, heteroaryl,cycloalkyl or heterocyclyl;

wherein R^(Y′) is selected from hydrido, alkyl, alkenyl, alkynyl, aryl,heteroaryl, cycloalkyl, heterocyclyl or hydroxyl;

wherein A′ is H, NH₂, NHR^(A′), NR^(A′)R^(B′), alkyl, alkenyl, alkynyl,alkoxy, aryloxy, aryl, heteroaryl, cycloalkyl or heterocyclyl;

wherein R^(A′) and R^(B′) are independently selected from alkyl,alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, heterocyclyl orcarboalkoxy;

wherein when m is 0, then A′ is additionally selected from:

wherein each of R⁵⁰-R⁵³ is independently selected from C₁-C₁₅ alkyl,alternatively, wherein B′ and A′ together form a 5-7 memberedheterocyclic or heteroaryl ring.

Wherein R² is

wherein K and K′ together form a C₃-C₇ cycloalkyl or heterocyclyl ringor a C₅-C₁₀ aryl or heteroaryl ring;

wherein J is selected from the group consisting of hydrido, amino,NHR^(J), NR^(J)R^(K), alkyl, alkenyl, alkynyl, alkoxy, aryloxy, aryl,heteroaryl, cycloalkyl, heterocyclyl, alkylamino, hydroxyl, thio,alkylthio, alkenylthio, sulfinyl, sulfonyl, azido, cyano, halo,

wherein each of R²⁴, R²⁵, and R²⁶ is independently selected from thegroup consisting of alkyl, cycloalkyl, heterocyclyl, aryl andheteroaryl; or R²⁴ and R²⁵ together form a 5-8 membered heterocyclylring;

wherein R^(J) and R^(K) are independently selected from alkyl, alkenyl,alkynyl, aryl, heteroaryl, cycloalkyl or heterocyclyl; or

alternatively, wherein J, together with R¹⁷, forms a 5-8 memberedheterocyclyl or cycloalkyl ring; or

alternatively, wherein J, together with both R¹⁷ and R¹⁸, forms a 5-8membered aryl, cycloalkyl, heterocyclyl or heteroaryl ring; and

wherein each of R¹⁷ and R¹⁸ is independently selected from the groupconsisting of hydrido, hydroxyl, halo, alkoxy, amino, thio, sulfinyl,sulfonyl and

or

wherein R¹⁷ and R¹⁸ taken together can form a group consisting of ketal,thioketal,

wherein each of R²² and R²³ is independently selected from the groupconsisting of hydrido and alkyl.

In another embodiment, the invention also provides pharmaceuticalcompositions comprising compounds of Formula I and methods of usethereof.

In a further embodiment, the invention provides methods of makingcompounds of Formula I and pharmaceutical compositions thereof.

In an even further embodiment, the invention provides compounds usefulas intermediates for the preparation of the compounds of Formula I

In a still further embodiment, the invention provides methods of use ofthe compounds of Formula I to treat bacterial infections in humans

DETAILED DESCRIPTION OF THE INVENTION

Definitions

Molecular terms, when used in this application, have their commonmeaning unless otherwise specified.

The term “hydrido” denotes a single hydrogen atom (H)

The term “acyl” is defined as a carbonyl radical attached to an alkyl,alkenyl, alkynyl, cycloalkyl, heterocycyl, aryl or heteroaryl group,examples including, without limitation, such radicals as acetyl andbenzoyl.

The term “amino” denotes a nitrogen radical containing two substituentsindependently selected from the group consisting of hydrido, alkyl,cycloalkyl, carboalkoxy, heterocyclyl, aryl, heteroaryl and sulfonyl.Subsets of the term amino are (1) the term “unsubstituted amino” whichdenotes an NH₂ radical, (2) the term “mono substituted amino” which isdefined as a nitrogen radical containing a hydrido group and asubstituent group selected from alkyl, cycloalkyl, heterocyclyl, aryl,or heteroaryl, and (3) the term “disubstituted amino” which is definedas a nitrogen radical containing two substituent groups independentlyselected from, alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl.Preferred mono substituted amino radicals are “lower mono substitutedamino” radicals, whereby the substituent group is a lower alkyl group.Preferred disubstituted amino radicals are “lower disubstituted amino”radicals, whereby the substituent groups are lower alkyl.

The term “acyloxy” denotes an oxygen radical adjacent to an acyl group.

The term “acylamino” denotes a nitrogen radical adjacent to an acylgroup.

The term “carboalkoxy” is defined as a carbonyl radical adjacent to analkoxy or aryloxy group.

The term “carboxyamido” denotes a carbonyl radical adjacent to an aminogroup.

The term “halo” is defined as a bromo, chloro, fluoro or iodo radical.

The term “thio” denotes a radical containing a substituent groupindependently selected from hydrido, alkyl, cycloalkyl, heterocyclyl,aryl and heteroaryl, attached to a divalent sulfur atom, such as,methylthio and phenylthio.

The term “alkyl” is defined as a linear or branched, saturated radicalhaving one to about twenty carbon atoms unless otherwise specified.Preferred alkyl radicals are “lower alkyl” radicals having one to aboutfive carbon atoms. One or more hydrogen atoms can also be replaced by asubstitutent group selected from acyl, amino, acylamino, acyloxy,carboalkoxy, carboxy, carboxyamido, cyano, halo, hydroxyl, nitro, thio,alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl,alkoxy, aryloxy, sulfinyl, sulfonyl, oxo, guanidino, formyl and an aminoacid side chain. Examples of alkyl groups include, without limitation,methyl, tertbutyl, isopropyl, and methoxymethyl. Subsets of the termalkyl are (1) “unsubstituted alkyl” which is defined as an alkyl groupthat bears no substituent groups (2) “substituted alkyl” which denotesan alkyl radical in which (a) one or more hydrogen atoms is replaced bya substitutent group selected from acyl, acyloxy, carboalkoxy, carboxy,carboxyamido, cyano, nitro, thio, alkyl, alkenyl, alkynyl, cycloalkyl,heterocyclyl, aryl, heteroaryl, alkoxy, aryloxy, sulfinyl, sulfonyl,N-sulfonylcarboxyamido, N-acylaminosulfonyl or (b) two or more hydrogenatoms are each replaced by a substituent group independently selectedfrom hydroxyl, carboxy, C₁-C₃ alkoxy, amino, acylamino, oxo orguanidino; and (3) the term “selected substituted alkyl” which denotesan alkyl radical in which (a) one proton is replaced by a group selectedfrom hydroxyl, carboxy C₁-C₃ alkoxy, unsubstituted amino, acylamino, oracylamino phenyl or (b) one to three protons is replaced by a halosubstituent.

The term “alkenyl” is defined as linear or branched radicals having twoto about twenty carbon atoms, preferably three to about ten carbonatoms, and containing at least one carboncarbon double bond. One or morehydrogen atoms can also be replaced by a substituent group selected fromacyl, amino, acylamino, acyloxy, carboalkoxy, carboxy, carboxyamido,cyano, halo, hydroxyl, nitro, thio, alkyl, alkenyl, alkynyl, cycloalkyl,heterocyclyl, aryl, heteroaryl, alkoxy, aryloxy, sulfinyl, sulfonyl,formyl, oxo and guanidino. The double bond portion(s) of the unsaturatedhydrocarbon chain may be either in the cis or trans configuration.Examples of alkenyl groups include, without limitation, ethylenyl orphenyl ethylenyl.

The term “alkynyl” denotes linear or branched radicals having from twoto about ten carbon atoms, and containing at least one carbon-carbontriple bond One or more hydrogen atoms can also be replaced by asubstituent group selected from acyl, amino, acylamino, acyloxy,carboalkoxy, carboxy, carboxyamido, cyano, halo, hydroxyl, nitro, thio,alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl,alkoxy, aryloxy, sulfinyl, sulfonyl, formyl, oxo and guanidino. Anexample of alkynyl group includes, without limitation, propynyl.

The term “aryl” or “aryl ring” denotes aromatic radicals in a single orfused carbocyclic ring system, having from five to fourteen ringmembers. In a preferred embodiment, the ring system has from six to tenring members. One or more hydrogen atoms may also be replaced by asubstituent group selected from acyl, amino, acylamino, acyloxy, azido,alkylthio, carboalkoxy, carboxy, carboxyamido, cyano, halo, hydroxyl,nitro, thio, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl,heteroaryl, alkoxy, aryloxy, sulfinyl, sulfonyl and formyl. Examples ofaryl groups include, without limitation, phenyl, naphthyl, biphenyl,terphenyl. Subsets of the term aryl are (1) the term “phenyl” whichdenotes a compound of the formula.

(2) the term “substituted phenyl” which is defined as a phenyl radicalin which one or more protons are replaced by a substituent groupselected from acyl, amino, acyloxy, azido, alkylthio, carboalkoxy,carboxy, carboxyamido, cyano, halo, hydroxyl, nitro, thio, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, alkoxy,aryloxy, sulfinyl, sulfonyl, N-sulfonylcarboxyamido, andN-acylaminosulfonyl and (3) the term “acylamino phenyl” denotes a phenylradical in which one hydrogen atom is replaced by an acylamino group.One or more additional hydrogen atoms can also be replaced by asubstituent group selected from acyl, amino, acylamino, acyloxy, azido,alkylthio, carboalkoxy, carboxy, carboxyamido, cyano, halo, hydroxyl,nitro, thio, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl,heteroaryl, alkoxy, aryloxy, sulfinyl, sulfonyl, N-sulfonylcarboxyamido,and N-acylaminosulfonyl.

“Heteroaryl” or “heteroaryl ring” denotes an aromatic radical whichcontain one to four hetero atoms or hetero groups selected from O, N, S,

in a single or fused heterocyclic ring system, having from five tofifteen ring members. In a preferred embodiment, the heteroaryl ringsystem has from six to ten ring members, One or more hydrogen atoms mayalso be replaced by a substituent group selected from acyl, amino,acylamino, acyloxy, carboalkoxy, carboxy, carboxyamido, cyano, halo,hydroxyl, nitro, thio, thiocarbonyl, alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclyl, aryl, heteroaryl, alkoxy, aryloxy, sulfinyl,sulfonyl, and formyl. Examples of heteroaryl groups include, withoutlimitation, pyridinyl, thiazolyl, thiadiazoyl, isoquinolinyl, pyrazolyl,oxazolyl, oxadiazoyl, triazolyl, and pyrrolyl groups. Subsets of theterm heteroaryl are (1) the term “pyridinyl” which denotes compounds ofthe formula:

(2) the term “substituted pyridinyl” which is defined as a pyridinylradical in which one or more protons is replaced by a substituent groupselected from acyl, amino, acyloxy, carboalkoxy, carboxy, carboxyamido,cyano, halo, hydroxyl, nitro, thio, alkyl, alkenyl, alkynyl, cycloalkyl,heterocyclyl, aryl, heteroaryl, alkoxy, aryloxy, sulfinyl, sulfonyl,N-sulfonylcarboxyamido, and N-acylaminosulfonyl and (3) the term“acylamino pyridinyl” which denotes a pyridinyl radical in which onehydrogen atom is replaced by an acylamino group, additionally, one ormore additional hydrogen atoms can also be replaced by a substituentgroup selected from acyl, amino, acylamino, acyloxy, carboalkoxy,carboxy, carboxyamido, cyano, halo, hydroxyl, nitro, thio, thiocarbonyl,alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl,alkoxy, aryloxy, sulfinyl, sulfonyl, N-sulfonylcarboxyamido, andN-acylaminosulfonyl.

The term “cycloalkyl” or “cycloalkyl ring” is defined as a saturated orpartially unsaturated carbocyclic ring in a single or fused carbocyclicring system having from three to twelve ring members. In a preferredembodiment, a cycloalkyl is a ring system having three to seven ringmembers. One or more hydrogen atoms may also be replaced by asubstituent group selected from acyl, amino, acylamino, acyloxy,carboalkoxy, carboxy, carboxyamido, cyano, halo, hydroxyl, nitro, thio,alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl,alkoxy, aryloxy, sulfinyl, sulfonyl and formyl. Examples of a cycloalkylgroup include, without limitation, cyclopropyl, cyclobutyl, cyclohexyl,and cycloheptyl.

The term “heterocyclyl,” “heterocyclic” or “heterocyclyl ring” isdefined as a saturated or partially unsaturated ring containing one tofour hetero atomsor hetero groups selected from O, N, NH,

wherein R^(Z) is as defined for R^(X),

in a single or fused heterocyclic ring system having from three totwelve ring members. In a preferred embodiment, a heterocyclyl is a ringsystem having three to seven ring members One or more hydrogen atoms mayalso be replaced by a substituent group selected from acyl, amino,acylamino, acyloxy, oxo, thiocarbonyl, imino, carboalkoxy, carboxy,carboxyamido, cyano, halo, hydroxyl, nitro, thio, alkyl, alkenyl,alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, alkoxy, aryloxy,sulfinyl, sulfonyl and formyl Examples of a heterocyclyl group include,without limitation, morpholinyl, piperidinyl, and pyrrolidinyl.

The term “alkoxy” denotes oxy-containing radicals substituted with analkyl, cycloalkyl or heterocyclyl group. Examples include, withoutlimitation, methoxy, tert-butoxy, benzyloxy and cyclohexyloxy.

The term “aryloxy” denotes oxycontaining radicals substituted with anaryl or heteroaryl group. Examples include, without limitation, phenoxy.

The term “amino acid side chain” denotes any side chain (R group) from anaturally-occurring or a non-naturally occurring amino acid.

The term “sulfinyl” is defined as a tetravalent sulfur radicalsubstituted with an oxo substituent and a second substituent selectedfrom the group consisting of alkyl, cycloalkyl, heterocyclyl, aryl, orheteroaryl group.

The term “sulfonyl” is defined as a hexavalent sulfur radicalsubstituted with two oxo substituents and a third substituent selectedfrom alkyl, cycloalkyl, heterocyclyl aryl, or heteroaryl.

The term “carbamate amino protecting group” is defined as a recognizedamino protecting group that when bound to an amino group forms acarbamate. Examples of carbamate amino protecting groups can be found in“Protective Groups in Organic Synthesis” by Theodora W. Greene, JohnWiley and Sons, New York, 1981. Examples of carbamate amino protectinggroups include benzyloxycarbonyl, t-butoxycarbonyl, t-amyloxycarbonyl,isobornyloxycarbonyl, adamantyloxycarbonyl, chlorobenzyloxycarbonyl,nitrobenzyloxycarbonyl or the like.

The salts of the compounds of the invention (preferably a compound ofFormula I) include acid addition salts and base addition salts. In apreferred embodiment, the salt is a pharmaceutically acceptable salt ofthe compound of Formula I. The term “pharmaceutically-acceptable salts”embraces salts commonly used to form alkali metal salts and to formaddition salts of free acids or free bases The nature of the salt is notcritical, provided that it is pharmaceutically-acceptable. Suitablepharmaceutically-acceptable acid addition salts of the compounds of theinvention (preferably a compound of Formula I) may be prepared from aninorganic acid or an organic acid. Examples of such inorganic acidsinclude, without limitation, hydrochloric, hydrobromic, hydroiodic,nitric, carbonic, sulfuric and phosphoric acid. Appropriate organicacids may be selected from aliphatic, cycloaliphatic, aromatic,arylaliphatic, heterocyclic, carboxylic and sulfonic classes of organicacids, examples of which include, without limitation, formic, acetic,propionic, succinic, glycolic, gluconic, maleic, embonic (pamoic),methanesulfonic, ethanesulfonic, 2-hydroxyethanesulfonic, pantothenic,benzenesulfonic, toluenesulfonic, sulfanilic, mesylic,cyclohexylaminosulfonic, stearic, algenic, β-hydroxybutyric, malonic,galactic, and galacturonic acid. Suitable pharmaceutically-acceptablebase addition salts of compounds of the invention (preferably a compoundof Formula I) include, but are not limited to, metallic salts made fromaluminum, calcium, lithium, magnesium, potassium, sodium and zinc ororganic salts made from N,N′-dibenzylethylenediamine, chloroprocaine,choline, diethanolamine, ethylenediamine, N-methylglucamine, lysine andprocaine. All of these salts may be prepared by conventional means fromthe corresponding compound of the invention (preferably a compound ofFormula I) by treating, for example, the compound of the invention(preferably a compound of Formula I) with the appropriate acid or base.

The compounds of the invention (preferably compounds of Formula I) canpossess one or more asymmetric carbon atoms and are thus capable ofexisting in the form of optical isomers as well as in the form ofracemic or non-racemic mixtures thereof. The compounds of the invention(preferably compounds of Formula I) can be utilized in the presentinvention as a single isomer or as a mixture of stereochemical isomericforms. Diastereoisomers, i.e., nonsuperimposable stereochemical isomers,can be separated by conventional means such as chromatography,distillation, crystallization or sublimation. The optical isomers can beobtained by resolution of the racemic mixtures according to conventionalprocesses, for example by formation of diastereoisomeric salts bytreatment with an optically active acid or base. Examples of appropriateacids include, without limitation, tartaric, diacetyltartaric,dibenzoyltartaric, ditoluoyltartaric and camphorsulfonic acid. Themixture of diastereomers can be separated by crystallization followed byliberation of the optically active bases from these salts. Analternative process for separation of optical isomers includes the useof a chiral chromatography column optimally chosen to maximize theseparation of the enantiomers. Still another available method involvessynthesis of covalent diastereoisomeric molecules by reacting compoundsof the invention (preferably compounds of Formula I) with an opticallypure acid in an activated form or an optically pure isocyanate. Thesynthesized diastereoisomers can be separated by conventional means suchas chromatography, distillation, crystallization or sublimation, andthen hydrolyzed to obtain the enantiomerically pure compound. Theoptically active compounds of the invention (preferably compounds ofFormula I) can likewise be obtained by utilizing optically activestarting materials. These isomers may be in the form of a free acid, afree base, an ester or a salt.

The invention also embraces isolated compounds. An isolated compoundrefers to a compound which represents at least 10%, preferably at least20%, more preferably at least 50% and most preferably at least 80% ofthe compound present in the mixture. In a preferred embodiment, thecompound, a pharmaceutically acceptable salt thereof, or apharmaceutical composition comprising the compound exhibits a detectable(i.e. statistically significant) antimicrobial activity when tested inconventional biological assays such as those described herein.

Lipopeptide Compounds

The invention provides a compound of formula (I).

wherein X and X″ are independently selected from C═O, C═S, C═NH,C═NR^(X), S═O or SO₂;

wherein n is 0 or 1;

wherein R^(X) is selected from alkyl, alkenyl, alkynyl, aryl,heteroaryl, cycloalkyl, heterocyclyl, hydroxyl, alkoxy, carboxy orcarboalkoxy;

wherein B is X″R^(Y), H, alkyl, alkenyl, alkynyl, aryl, heteroaryl,cycloalkyl or heterocyclyl; and

wherein R^(Y) is selected from hydrido, alkyl, alkenyl, alkynyl, aryl,heteroaryl, cycloalkyl, heterocyclyl or hydroxyl.

In one aspect, A is H, NH₂, NHR^(A), NR^(A)R^(B), heteroaryl, cycloalkylor heterocyclyl;

wherein R^(A) and R^(B) are independently selected from alkyl, alkenyl,alkynyl, aryl, heteroaryl, cycloalkyl, heterocyclyl or carboalkoxy;

wherein when n is 0, then A is additionally selected from:

wherein each R⁵⁰-R⁵³ is independently selected from (C₁-C₁₅) alkyl,

provided that when B is H and X is C═O, then A is other than

(a) a pyridinyl ring substituted with a single NHC(O)R^(D) substitutentor

(b) a (C₅-C₆) saturated cycloalkyl ring substituted with a singleNHC(O)R^(D) substitutent, wherein R^(D) is (C₁-C₁₇) unsubstituted alkylor (C₂-C₁₇) unsubstituted alkenyl; and

when B is H and n is 0, then A is not H.

In another aspect, A is aryl;

provided that when B is H and X is C═O, then A is other than a phenylring substituted with either:

(a) —O—((C₈-C₁₅) unsubstituted alkyl), wherein said phenyl ring may befurther optionally substituted with one substituent selected from halo,nitro, (C₁-C₃) alkyl, hydroxyl, (C₁-C₃) alkoxy or (C₁-C₃) alkylthio; or

(b) —NHC(O)R^(D), wherein the phenyl ring may be further optionallysubstituted with 1-2 substituents independently selected from amino,nitro, (C₁-C₃) alkyl, hydroxyl, (C₁-C₃) alkoxy, halo, mercapto, (C₁-C₃)alkylthio, carbamyl or (C₁-C₃) alkylcarbamyl; wherein R^(D) is asdefined previously.

In a third aspect of the invention, A is alkyl, alkenyl, alkynyl, alkoxyor aryloxy;

provided that when B is H and X is C═O, then A is other than

(a) —(C₁-C₁₆ unsubstituted alkyl)-NH₂;

(b) —(C₁-C₁₀ unsubstituted alkyl)-NHC(O)R^(D), wherein R^(D) is asdefined previously;

(c) —(C₁-C₁₈)-alkyl, optionally substituted with up to one hydroxyl,carboxyl or C₁-C₃ alkoxy, or one to three halo substituents;

(d) —(C₄-C₁₈)-unsubstituted alkenyl;

wherein R⁵⁴ is selected from C₁-C₁₇-unsubstituted alkyl orC₂-C₁₇-unsubstituted alkenyl; wherein R⁵⁵ is selected from hydroxyethyl,hydroxymethyl, mercaptomethyl, mercaptoethyl, methylthioethyl,2-thienyl, 3-indolemethyl, phenyl optionally substituted with a groupselected from halo, nitro, C₁-C₃-unsubstituted alkyl, hydroxy,C₁-C₃-unsubstituted alkoxy, C₁-C₃-unsubsituted alkylthio, carbamyl orC₁-C₃-unsubstituted alkylcarbamyl; or benzyl optionally substituted witha group selected from halo, nitro, C₁-C₃-unsubstituted alkyl, hydroxy,C₁-C₃-unsubstituted alkoxy, C₁-C₃-unsubsituted alkylthio, carbamyl orC₁-C₃ unsubstituted alkylcarbamyl; wherein t is 0 or 1 and wherein u isan integer from 1-3; and

when B is H and X is C═O, then X, together with A, does not form acarbamate amino protecting group; and

when B is H and n is 0, then A is other than C₄-C₁₄ unsubstituted alkyl.

In a fourth aspect, B and A together form a 5-7 membered heterocyclic orheteroaryl ring.

Wherein R¹ is

wherein X′ and X′″ are independently selected from C═O, C═S, C═NH,C═NR^(X′), S═O or SO₂;

wherein m is 0 or 1;

wherein R^(X′) is selected from alkyl, alkenyl, alkynyl, aryl,heteroaryl, cycloalkyl, heterocyclyl, hydroxyl, alkoxy, carboxy orcarboalkoxy;

wherein B′ is X′″ R^(Y′), H, alkyl, alkenyl, alkynyl, aryl, heteroaryl,cycloalkyl or heterocyclyl;

wherein R^(y) is selected from hydrido, alkyl, alkenyl, alkynyl, aryl,heteroaryl, cycloalkyl, heterocyclyl or hydroxyl;

wherein A′ is H, NH₂, NHR^(A′), NR^(A′)R^(B′), alkyl, alkenyl, alkynyl,alkoxy, aryloxy, aryl, heteroaryl, cycloalkyl or heterocyclyl;

wherein R^(A′) and R^(B′) are independently selected from alkyl,alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, heterocyclyl orcarboalkoxy;

wherein when m is 0, then A′ is additionally selected from

wherein each of R⁵⁰-R⁵³ is independently selected from C₁-C₁₅ alkyl;

alternatively, wherein B′ and A′ together form a 5-7 memberedheterocyclic or heteroaryl ring.

Wherein R² is

wherein K and K′ together form a C₃-C₇ cycloalkyl or heterocyclyl ringor a C₅-C₁₀ aryl or heteroaryl ring;

wherein J is selected from the group consisting of hydrido, amino,NHR^(J), NR^(J)R^(K), alkyl, alkenyl, alkynyl, alkoxy, aryloxy, aryl,heteroaryl, cycloalkyl, heterocyclyl, alkylamino, hydroxyl, thio,alkylthio, alkenylthio, sulfinyl, sulfonyl, azido, cyano, halo,

wherein each of R²⁴, R²⁵, and R²⁶ is independently selected from thegroup consisting of alkyl, cycloalkyl, heterocyclyl, aryl andheteroaryl; or R²⁴ and R²⁵ together form a 5-8 membered heterocyclylring;

wherein R^(J) and R^(K) are independently selected from alkyl, alkenyl,alkynyl, aryl, heteroaryl, cycloalkyl or heterocyclyl; or

alternatively, wherein J, together with R¹⁷, forms a 5-8 memberedheterocyclyl or cycloalkyl ring; or

alternatively, wherein J, together with both R¹⁷ and R¹⁸, forms a 5-8membered aryl, cycloalkyl, heterocyclyl or heteroaryl ring; and

wherein each of R¹⁷ and R¹⁸ is independently selected from the groupconsisting of hydrido, halo, hydroxyl, alkoxy, amino, thio, sulfinyl,sulfonyl and

or

wherein R¹⁷ and R¹⁸ taken together can form a group consisting of ketal,thioketal,

wherein each of R²² and R²³ is independently selected from the groupconsisting of hydrido and alkyl

In a preferred embodiment of the invention, R is selected from

wherein each of R³, R⁴ R⁵, and R⁶ is independently selected from thegroup consisting of hydrido, alkyl, aryl, heterocyclyl and heteroaryl,and wherein R⁴⁴ is selected from the group consisting of alkyl, aryl,heterocyclyl and heteroaryl.

In a more preferred embodiment of the invention R is selected from

wherein R^(4′) is selected from the group consisting of alkyl,substituted alkyl, substituted phenyl, heteroaryl, heterocyclyl,optionally substituted (C₈-C₁₄)straight chain alkyl and

wherein R⁷ is an alkyl group.

In an even more preferred embodiment of the invention, R is

wherein X³ is chloro or trifluoromethyl and wherein q is 0 or 1.

In a preferred embodiment of the invention, R¹ is selected from thegroup consisting of:

wherein R⁸ is selected from an amino acid side chain, wherein said aminoacid side chain may be one that is naturally occurring or one that isnot naturally occurring, wherein each of R⁹, R¹⁰ and R¹¹ is selectedfrom hydrido, alkyl, aryl, heterocyclyl and heteroaryl; wherein R¹² isselected from the group consisiting of heterocyclyl, heteroaryl, aryl,and alkyl and wherein R¹³ is selected from (C₁-C₃)alkyl and aryl.

In a more preferred embodiment of the invention, R¹ is selected from thegroup consisting of

wherein R⁸ is selected from tryptophan side chain and lysine side chain;wherein each of R¹⁰ and R¹¹ is independently selected from hydrido andalkyl; wherein R¹² is selected from imidazolyl, N-methylimidazolyl,indolyl, quinolinyl, benzyloxybenzyl, and benzylpiperidenylbenzyl; andwherein X⁴ is selected from fluoro and trifluoromethyl.

In a preferred embodiment of R², J is selected from the group consistingof hydrido, amino, azido and

wherein R¹⁷ and R¹⁸ taken together form a group selected from the groupconsisting of ketal,

alternatively, R¹⁷ is hydroxyl when R¹⁸ is hydrido. Alternatively,wherein J, together with R¹⁷, forms a heterocyclyl ring.

In a more preferred embodiment of the invention, R² is selected from

wherein R¹⁷ and R¹⁸ taken together form a group selected from

wherein R²² is selected from the group consisting of H and alkyl;wherein R¹⁹ is selected from the group consisting of hydrido, amino,azido and

In an even more preferred embodiment of the invention R² is

Table I provides exemplary compounds of Formula I:

TABLE I Cpd Mass Synth # R R¹ R² Spec Ex # 1 NHCONH(CH₂)₇CH₃ NH₂

1622.8 1 2 NHCONH(CH₂)₁₁CH₃ NH₂

1665 2 3 NHCONH(CH₂)₁₀CH₃

1951 3 5

1867 3 6

1935 3 7 NH(CH₂)₈CH₃

1779  3a 8 NHCO(CH₂)₈CO₂CH₃

1851 3 9 NHCO(CH₂)₆CO₂CH₃

1823 3 10 NHCO(CH₂)₆NHBoc

1980 3 11 NHCO(CH₂)₇NHBoc

1894 3 12 NHCO(CH₂)₁₀NHBoc

1936 3 13 NHCO(CH₂)₁₁NHBoc

1950 3 17 NHCONH(CH₂)₁₁CH₃

1865  3b 18

NH₂

1696  1a 19

NH₂

1668 1 20

1807 3 21

1841 3 22

1864 3 23

1843 3 24

1882 3 25

1823.3 4 34

1738 3 35

1862 3 36

1962 3 40

NH₂

1736 1 41

NHBoc

1836 1 43

NHBoc

1624 1 44

NHBoc

1675 1 48 NHCONH(CH₂)₁₀CH₃ NH₂

1665  2a 49

NH₂

1703 1 50

1738.8 3 56 NHCONH(CH₂)₇CH₃

1950 4 57 NHCONH(CH₂)₁₀CH₃

1992 4 58 NHCONH(CH₂)₁₁CH₃

2006 4 62 NHCONH(CH₂)₇CH₃

1750 4 63 NHCONH(CH₂)₁₀CH₃

1792 4 64 NHCONH(CH₂)₁₁CH₃

1806 4 69 NHCONH(CH₂)₇CH₃

1808 4 70 NHCONH(CH₂)₇CH₃

1759 4 71 NHCONH(CH₂)₇CH₃

1650 3 75 NHCONH(CH₂)₁₀CH₃

1706.9 3 76 NHCONH(CH₂)₇CH₃

1780.9  4a 77 NHCONH(CH₂)₇CH₃

1701.8  4a 78 NHCONH(CH₂)₇CH₃

1807.9  4a 87 NHCONH(CH₂)₁₁CH₃

1757.9  4a 88 NHCONH(CH₂)₁₁CH₃

1864  4a 89 NHCONH(CH₂)₁₁CH₃

1837  4a 100

NH₂

1635.7 1 106

1832 4 108 NHCONH(CH₂)₁₀CH₃

1801 4 113 NHCONH(CH₂)₁₀CH₃

1743  4a 114 NHCONH(CH₂)₁₀CH₃

1822  4b 115

NHBoc

1828.8 1 116

NH₂

1729 1 117 NHCONH(CH₂)₈CH₃ NHBoc

1636.6  2b 118 NHCONH(CH₂)₈CH₃ NH₂

1636.6  2b 119 NHCONH(CH₂)₉CH₃ NHBoc

1650.1  2c 120 NHCONH(CH₂)₉CH₃ NH₂

1650.2  2c 123 NHCOCH₂S(CH₂)₁₁CH₃ NH₂

1709 1 124 NHCOCH₂S(CH₂)₁₀CH₃ NH₂

1695 1 125 NHCOCH₂S(CH₂)₉CH₃ NH₂

1681 1

Preferred compounds of Formula I are composed 2, compound 3, compound18, compound 48, compound 89, compound 116, compund 118, and compound120.

Other preferred compounds include a compound of Formula (I′).

wherein R¹⁰⁰, R¹⁰¹ and R¹⁰² are as defined in Table II.

TABLE II Cpd Mass Synth # R¹⁰⁰ R¹⁰¹ R¹⁰² Spec Ex # 72

NHBoc

1764.5 1 73

NHBoc

1792.5 1 74

NHBoc

1820.5 1 109 NHCOCHCH(CH₂)₇CH₃ NHBoc

1651.8  1b 110 NHCOCHCH(CH₂)₉CH₃ NHBoc

1679.9  1b 111 NHCOCHCH(CH₂)₇CH₃ NH₂

1680  1b 112 NHCOCHCH(CH₂)₉CH₃ NH₂

1680  1b

According to a preferred embodiment, the present invention provides oneor more crystalline forms of compound of formula (I), and salts thereof.

Lipopeptide Intermediates

The present invention also provides compounds that are particularlyuseful as intermediates for the preparation of the compounds of FormulaI. These compounds may also have antibacterial properties, as discussedabove. In one aspect of the invention, compounds of Formula II areprovided:

wherein R¹⁴ is selected from the group consisting of

wherein R⁵⁶ is an optionally substituted straight-chain C₈-C₁₄ alkylgroup and wherein q′ is 0-3.

Compounds 1, 2, 18, 48, 116, 118 and 120 are useful both asantibacterial compounds and as intermediates in the synthesis ofcompounds of this invention.

Compounds 72, 73 and 74 as well as the formula (II) compounds in TableIII are other preferred compounds that are useful as antibacterialcompounds and as intermediates in the synthesis of compounds of thisinvention:

TABLE III Compound # R¹⁴ 45

37

46

38

47

39

Table IV provides another set of formula (II) compounds that are usefulas intermediates in the synthesis of compounds of this invention:

TABLE IV Compound # R¹⁴ 150 (CH₂)₇CH₃ 151 (CH₂)₈CH₃ 152 (CH₂)₉CH₃ 153(CH₂)₁₀CH₃ 154 (CH₂)₁₁CH₃ 155 (CH₂)₁₂CH₃

In another aspect of the invention, compounds of Formula III areprovided as useful intermediates for the preparation of compounds ofFormula I and/or antibacterial compounds:

wherein R¹⁵ is selected from hydrido and a carbamate amino protectinggroup, preferably a tert-butoxycarbonyl group; wherein R¹⁶ is selectedfrom the group consisting of

wherein R⁵⁷ is a halo or halo substituted alkyl group, preferably afluoro or trifluoromethyl group; wherein, R²⁰ is an amino acid sidechain, preferably a lysine or tryptophan side chain.Lipopeptide Compound Pharmaceutical Compositions and Methods of Usethereof

Another object of the instant invention is to provide lipopeptidecompounds or salts thereof, as well as pharmaceutical compositions orformulations comprising lipopeptide compounds or its salts.

Lipopeptide compounds, or pharmaceutically acceptable salts thereof, canbe formulated for oral, intravenous, intramuscular, subcutaneous orparenteral administration for the therapeutic or prophylactic treatmentof diseases, particularly bacterial infections. For oral or parenteraladministration, lipopeptide compounds of this invention can be mixedwith conventional pharmaceutical carriers and excipients and used in theform of tablets, capsules, elixirs, suspensions, syrups, wafers and thelike. The compositions comprising a compound of this invention willcontain from about 0.1 to about 99% by weight of the active compound,and more generally from about 10 to about 30%.

The pharmaceutical preparations disclosed herein are prepared inaccordance with standard procedures and are administered at dosages thatare selected to reduce, prevent or eliminate the infection (See, e. g.,Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton,Pa. and Goodman and Gilman's. The Pharmaceutical Basis of Therapeutics,Pergamon Press, New York, N.Y., the contents of which are incorporatedherein by reference, for a general description of the methods foradministering various antimicrobial agents for human therapy). Thecompositions of the invention (preferably of Formula I) can be deliveredusing controlled (e.g., capsules) or sustained release delivery systems(e.g., bioerodable matrices). Exemplary delayed release delivery systemsfor drug delivery that are suitable for administration of thecompositions of the invention (preferably of Formula I) are described inU.S. Pat. Nos. 4,452,775 (issued to Kent), 5,239,660 (issued toLeonard), 3,854,480 (issued to Zaffaroni).

The pharmaceuticallyacceptable compositions of the present inventioncomprise one or more compounds of the invention (preferably compounds ofFormula I) in association with one or more non-toxic,pharmaceuticallyacceptable carriers and/or diluents and/or adjuvantsand/or excipients, collectively referred to herein as “carrier”materials, and if desired other active ingredients. The compositions maycontain common carriers and excipients, such as corn starch or gelatin,lactose, sucrose, microcrystalline cellulose, kaolin, mannitol,dicalcium phosphate, sodium chloride and alginic acid. The compositionsmay contain croscarmellose sodium, microcrystalline cellulose, cornstarch, sodium starch glycolate and alginic acid.

Tablet binders that can be included are acacia, methylcellulose, sodiumcarboxymethylcellulose, polyvinylpyrrolidone (Povidone), hydroxypropylmethylcellulose, sucrose, starch and ethylcellulose.

Lubricants that can be used include magnesium stearate or other metallicstearates, stearic acid, silicone fluid, talc, waxes, oils and colloidalsilica.

Flavoring agents such as peppermint, oil of wintergreen, cherryflavoring or the like can also be used. It may also be desirable to adda coloring agent to make the dosage form more aesthetic in appearance orto help identify the product.

For oral use, solid formulations such as tablets and capsules areparticularly useful. Sustained release or enterically coatedpreparations may also be devised. For pediatric and geriatricapplications, suspensions, syrups and chewable tablets are especiallysuitable. For oral administration, the pharmaceutical compositions arein the form of, for example, a tablet, capsule, suspension or liquid.The pharmaceutical composition is preferably made in the form of adosage unit containing a therapeuticallyeffective amount of the activeingredient. Examples of such dosage units are tablets and capsules. Fortherapeutic purposes, the tablets and capsules which can contain, inaddition to the active ingredient, conventional carriers such as bindingagents, for example, acacia gum, gelatin, polyvinylpyrrolidone,sorbitol, or tragacanth; fillers, for example, calcium phosphate,glycine, lactose, maize-starch, sorbitol, or sucrose; lubricants, forexample, magnesium stearate, polyethylene glycol, silica, or talc;disintegrants, for example, potato starch, flavoring or coloring agents,or acceptable wetting agents. Oral liquid preparations generally are inthe form of aqueous or oily solutions, suspensions, emulsions, syrups orelixirs may contain conventional additives such as suspending agents,emulsifying agents, non-aqueous agents, preservatives, coloring agentsand flavoring agents. Examples of additives for liquid preparationsinclude acacia, almond oil, ethyl alcohol, fractionated coconut oil,gelatin, glucose syrup, glycerin, hydrogenated edible fats, lecithin,methyl cellulose, methyl or propyl parahydroxybenzoate, propyleneglycol, sorbitol, or sorbic acid.

For intravenous (IV) use, a lipopeptide compound according to theinvention can be dissolved or suspended in any of the commonly usedintravenous fluids and administered by infusion. Intravenous fluidsinclude, without limitation, physiological saline or Ringer's solution.Intravenous administration may be accomplished by using, withoutlimitation, syringe, minipump or intravenous line.

Formulations for parenteral administration can be in the form of aqueousor non-aqueous isotonic sterile injection solutions or suspensions.These solutions or suspensions can be prepared from sterile powders orgranules having one or more of the carriers mentioned for use in theformulations for oral administration. The compounds can be dissolved inpolyethylene glycol, propylene glycol, ethanol, corn oil, benzylalcohol, sodium chloride, and/or various buffers.

For intramuscular preparations, a sterile formulation of a lipopeptidecompound or a suitable soluble salt form of the compound, for examplethe hydrochloride salt, can be dissolved and administered in apharmaceutical diluent such as Water-for-Injection (WFI), physiologicalsaline or 5% glucose. A suitable insoluble form of the compound may beprepared and administered as a suspension in an aqueous base or apharmaceutically acceptable oil base, e.g., an ester of a long chainfatty acid such as ethyl oleate.

A dose of an intravenous, intramuscular or parental formulation of alipopeptide compound may be adminstered as a bolus or by slow infusion.A bolus is a dose that is administered in less than 30 minutes. In apreferred embodiment, a bolus is administered in less than 15 or lessthan 10 minutes. In a more preferred embodiment, a bolus is administeredin less than 5 minutes. In an even more preferred embodiment, a bolus isadministered in one minute or less. An infusion is a dose that isadministered at a rate of 30 minutes or greater. In a preferredembodiment, the infusion is one hour or greater. In another embodiment,the infusion is substantially constant.

For topical use the compounds of the present invention can also beprepared in suitable forms to be applied to the skin, or mucus membranesof the nose and throat, and can take the form of creams, ointments,liquid sprays or inhalants, lozenges, or throat paints. Such topicalformulations further can include chemical compounds such asdimethylsulfoxide (DMSO) to facilitate surface penetration of the activeingredient.

For application to the eyes or ears, the compounds of the presentinvention can be presented in liquid or semi-liquid form formulated inhydrophobic or hydrophilic bases as ointments, creams, lotions, paintsor powders.

For rectal administration the compounds of the present invention can beadministered in the form of suppositories admixed with conventionalcarriers such as cocoa butter, wax or other glyceride.

Alternatively, the compounds of the present invention can be in powderform for reconstitution in the appropriate pharmaceutically acceptablecarrier at the time of delivery. In another embodiment, the unit dosageform of the compound can be a solution of the compound or preferably asalt thereof in a suitable diluent in sterile, hermetically sealedampoules or sterile syringes. The concentration of the compound in theunit dosage may vary, e.g. from about 1 percent to about 50 percent,depending on the compound used and its solubility and the dose desiredby the physician. If the compositions contain dosage units, each dosageunit preferably contains from 1-500 mg of the active material. For adulthuman treatment, the dosage employed preferably ranges from 5 mg to 10g, per day, depending on the route and frequency of administration.

In another aspect, the invention provides a method for inhibiting thegrowth of microorganisms, preferably bacteria, comprising contactingsaid organisms with a compound of the invention, preferably a compoundof Formula I, under conditions which permit entry of the compound intosaid organism and into said microorganism. Such conditions are known toone skilled in the art and are exemplified in the Examples. This methodinvolves contacting a microbial cell with a therapeutically-effectiveamount of compound(s) of the invention, preferably compound(s) ofFormula I, in vivo or in vitro.

According to this aspect of the invention, the novel compositionsdisclosed herein are placed in a pharmaceutically acceptable carrier andare delivered to a recipient subject (preferably a human) in accordancewith known methods of drug delivery. In general, the methods of theinvention for delivering the compositions of the invention in vivoutilize art-recognized protocols for delivering the agent with the onlysubstantial procedural modification being the substitution of thecompounds of the invention (preferably compounds of Formula I) for thedrugs in the art-recognized protocols Likewise, the methods for usingthe claimed composition for treating cells in culture, for example, toeliminate or reduce the level of bacterial contamination of a cellculture, utilize artrecognized protocols for treating cell cultures withantibacterial agent(s) with the only substantial procedural modificationbeing the substitution of the compounds of the invention (preferablycompounds of Formula I) for the agents used in the art-recognizedprotocols.

In one embodiment, the invention provides a method for treating aninfection, especially those caused by gram-positive bacteria, in asubject with a therapeutically-effective amount of a lipopeptidecompound according to Formula I. Exemplary procedures for delivering anantibacterial agent are described in U.S. Pat No. 5,041,567, issued toRogers and in PCT patent application number EP94/02552 (publication noWO 95/05384), the entire contents of which documents are incorporated intheir entirety herein by reference. As used herein the phrase“therapeutically-effective amount” means an amount of a compound of thepresent invention that prevents the onset, alleviates the symptoms, orstops the progression of a bacterial infection. The term “treating” isdefined as administering, to a subject, a therapeutically-effectiveamount of a compound of the invention (preferably a compound of FormulaI) both to prevent the occurrence of an infection and to control oreliminate an infection. The term “subject”, as described herein, isdefined as a mammal, a plant or a cell culture. In a preferredembodiment, a subject is a human or other animal patient in need oflipopeptide compound treatment.

The method comprises administering to the subject an effective dose of acompound of this invention. An effective dose is generally between about0.1 and about 100 mg/kg of a lipopeptide compound of Formula I or apharmaceutically acceptable salt thereof. A preferred dose is from about0.1 to about 50 mg/kg of a lipopeptide compound of Formula I or apharmaceutically acceptable salt thereof. A more preferred dose is fromabout 1 to 25 mg/kg of a lipopeptide compound of Formula I or apharmaceutically acceptable salt thereof An effective dose for cellculture is usually between 0.1 and 1000 μg/mL, more preferably between0.1 and 200 μg/mL.

The compound of Formula I can be administered as a single daily dose orin multiple doses per day. The treatment regime may requireadministration over extended periods of time, e.g., for several days orfor from two to four weeks. The amount per administered dose or thetotal amount administered will depend on such factors as the nature andseverity of the infection, the age and general health of the patient,the tolerance of the patient to the compound and the microorganism ormicroorganisms involved in the infection. A method of administration toa patient of daptomycin, another member of the lipopeptide compoundclass is disclosed in U.S. Ser. No. 09/406,568, filed Sep. 24, 1999,which claims the benefit of U.S. Provisional Application Ser. Nos.60/101,828, filed Sep. 25, 1998, and 60/125,750, filed Mar. 24, 1999.

A lipopeptide compound according to this invention may also beadministered in the diet or feed of a patient or animal. If administeredas part of a total dietary intake, the amount of compound employed canbe less than 1% by weight of the diet and preferably no more than 0.5%by weight. The diet for animals can be normal foodstuffs to which thecompound can be added or it can be added to a premix.

The methods of the present invention comprise administering alipopeptide compound of Formula I or a pharmaceutical compositionthereof to a subject in need thereof in an amount that is efficacious inreducing or eliminating the bacterial infection. The compound may beadministered orally, parenterally, by inhalation, topically, rectally,nasally, buccally, vaginally, or by an implanted reservoir, externalpump or catheter. The compound may be prepared for opthalmic oraerosolized uses. The compounds of the present invention can beadministered as an aerosol for the treatment of pneumonia or otherlung-based infections. A preferred aerosol delivery vehicle is ananhydrous or dry powder inhaler. Lipopeptide compounds of Formula I or apharmaceutical composition thereof also may be directly injected oradministered into an abscess, ventricle or joint. Parenteraladministration includes subcutaneous, intravenous, intramuscular,intra-articular, intra-synovial, cisternal, intrathecal, intrahepatic,intralesional and intracranial injection or infusion. In a preferredembodiment, lipopeptide compounds are administered intravenously,subcutaneously or orally. In a preferred embodiment for administering alipopeptide compound according to Formula I to a cell culture, thecompound may be administered in a nutrient medium.

The method of the instant invention may be used to treat a subjecthaving a bacterial infection in which the infection is caused orexacerbated by any type of bacteria, particularly gram-positivebacteria. In one embodiment, a lipopeptide compound or a pharmaceuticalcomposition thereof is administered to a patient according to themethods of this invention. In a preferred embodiment, the bacterialinfection may be caused or exacerbated by gram-positive bacteria. Thesegram-positive bacteria include, but are not limited to,methicillin-susceptible and methicillin-resistant staphylococci(including Staphylococcus aureus, S. epidermidis, S. haemolyticus, S.hominis, S. saprophyticus, and coagulase-negative staphylococci),glycopeptide intermediary-susceptible S. aureus (GISA),penicillin-susceptible and penicillin-resistant streptococci (includingStreptococcus pneumoniae, S. pyogenes, S. agalactiae, S. avium, S.bovis, S. lactis, S. sangius and Streptococci Group C, StreptococciGroup G and viridans streptococci), enterococci (including vancomycinsusceptible and vancomycin-resistant strains such asEnterococcusfaecalis and E. faecium), Clostridium difficile, C.clostridiiforme, C. innocuum, C. perfringens, C. ramosum, Haemophilusinfluenzae, Listeria monocytogenes, Corynebacterium jeikeium,Bifidobacterium spp., Eubacterium aerofaciens, E. lentum, Lactobacillusacidophilus, L. casei, L. plantarum, Lactococcus spp., Leuconostoc spp.,Pediococcus, Peptostreptococcus anaerobius, P. asaccarolyticus, P.magnus, P. micros, P. prevotil, P. productus, Propionibacterium acnes,Actinomyces spp., Moraxella spp. (including M. catarrhalis) andEscherichia spp. (including E. coli).

In a preferred embodiment, the antibacterial activity of lipopeptidecompounds of Formula I against classically “resistant” strains iscomparable to that against classically “susceptible” strains in in vitroexperiments. In another preferred embodiment, the minimum inhibitoryconcentration (MIC) value for lipopeptide compounds according to thisinvention against susceptible strains is typically the same or lowerthan that of vancomycin. Thus, in a preferred embodiment, a lipopeptidecompound of this invention or a pharmaceutical composition thereof isadministered according to the methods of this invention to a patient whoexhibits a bacterial infection that is resistant to other compounds,including vancomycin or daptomycin. In addition, unlike glycopeptideantibiotics, lipopeptide compounds exhibits rapid,concentrationdependent bactericidal activity against gram-positiveorganisms. Thus, in a preferred embodiment, a lipopeptide compoundaccording to this invention or a pharmaceutical composition thereof isadministered according to the methods of this invention to a patient inneed of rapidly acting antibiotic therapy.

The method of the instant invention may be used for any bacterialinfection of any organ or tissue in the body. In a preferred embodiment,the bacterial infection is caused by gram-positive bacteria. Theseorgans or tissue include, without limitation, skeletal muscle, skin,bloodstream, kidneys, heart, lung and bone. The method of the inventionmay be used to treat, without limitation, skin and soft tissueinfections, bacteremia and urinary tract infections The method of theinvention may be used to treat community acquired respiratoryinfections, including, without limitation, otitis media, sinusitis,chronic bronchitis and pneumonia, including pneumonia caused bydrug-resistant S. pneumoniae or H. influenzae. The method of theinvention also may be used to treat mixed infections that comprisedifferent types of gram-positive bacteria, or which comprise bothgram-positive and gram-negative bacteria. These types of infectionsinclude intra-abdominal infections and obstetrical/gynecologicalinfections. The method of the invention also may be used to treat aninfection including, without limitation, endocarditis, nephritis, septicarthritis, intra-abdominal sepsis, bone and joint infections. andosteomyelitis. In a preferred embodiment, any of the above-describeddiseases may be treated using lipopeptide compounds according to thisinvention or pharmaceutical compositions thereof.

The method of the instant invention may also be practiced whileconcurrently administering one or more other antimicrobial agents, suchas antibacterial agents (antibiotics) or antifungal agents. In oneaspect, the method may be practiced by administering more than onelipopeptide compounds according to this invention. In anotherembodiment, the method may be practiced by administering a lipopeptidecompound according to this invention with another lipopeptide compound,such as daptomycin.

Antibacterial agents and classes thereof that may be co-administeredwith a compound of the present invention include, without limitation,penicillins and related drugs, carbapenems, cephalosporins and relateddrugs, aminoglycosides, bacitracin, gramicidin, mupirocin,chloramphenicol, thiamphenicol, fusidate sodium, lincomycin,clindamycin, macrolides, novobiocin, polymyxins, rifamycins,spectinomycin, tetracyclines, vancomycin, teicoplanin, streptogramins,anti-folate agents including sulfonamides, trimethoprim and itscombinations and pyrimethamine, synthetic antibacterials includingnitrofurans, methenamine mandelate and methenamine hippurate,nitroimidazoles, quinolones, fluoroquinolones, isoniazid, ethambutol,pyrazinamide, para-aminosalicylic acid (PAS), cycloserine, capreomycin,ethionamide, prothionamide, thiacetazone, viomycin, eveminomicin,glycopeptide, glycylcycline, ketolides, oxazolidinone; imipenen,amikacin, netilmicin, fosfomycin, gentamicin, ceftriaxone, Ziracin, LY333328, CL 331002, Linezolid, Synercid, Aztreonam, Metronidazole,Epiroprim, OCA-983, GV-143253, Sanfetrinem sodium, CS-834, Biapenem,A-99058.1, A-165600, A-179796, KA 159, Dynemicin A, DX8739, DU 6681;Cefluprenam, ER 35786, Cefoselis, Sanfetrinem celexetil, HGP-31,Cefpirome, HMR-3647, RU-59863, Mersacidin, KP 736, Rifalazil; Kosan, AM1732, MEN 10700, Lenapenem, BO 2502A, NE-1530, PR 39,(L-arginyl-L-ariginyl-L-arginyl-L-prolyl-L-arginyl-L-prolyl-L-prolyl-L-tryosyl-L-leucyl-L-prolyl-L-arginyl-L-prolyl-L-arginyl-L-prolyl-L-prolyl-L-prolyl-L-phenylalanyl-L-phenylalanyl-L-prolyl-L-prolyl-L-arginyl-L-leucyl-L-prolyl-L-prolyl-L-arginyl-L-isoleucyl-L-prolyl-L-prolylglycyl-L-phenylalanyl-L-prolyl-L-prolyl-L-arginyl-L-phenyalanyl-L-prolyl-L-prolyl-L-arginyl-L-phenylalanyl-L-prolinamide[SEQ. ID NO: 1]), K130, OPC 20000, OPC 2045, Veneprim, PD 138312, PD140248, CP 111905, Sulopenem, ritipenam acoxyl, RO-65-5788,Cyclothialidine, Sch-40832, SEP-132613, micacocidin A, SB-275833,SR-15402, SUN A0026, TOC 39, carumonam, Cefozopran, Cefetamct pivoxil,and T 3811.

In a preferred embodiment, antibacterial agents that may beco-administered with a compound according to this invention include,without limitation, imipenen, amikacin, netilmicin, fosfomycin,gentamicin, ceftriaxone, teicoplanin, Ziracin, LY 333328, CL 331002, R3647, Linezolid, Synercid, Aztreonam, and Metronidazole.

Antifungal agents that may be co-administered with a compound accordingto this invention include, without limitation, Caspofungen,Voriconazole, Sertaconazole, IB-367, FK-463, LY-303366, Sch-56592,Sitafloxacin, DB-289 polyenes, such as Amphotericin, Nystatin,Primaricin; azoles, such as Fluconazole, Itraconazole, and Ketoconazole;allylamines, such as Naftifine and Terbinafine; and anti-metabolitessuch as Flucytosine. Other antifungal agents include without limitation,those disclosed in Fostel et al., Drug Discovery Today 5:25-32 (2000),herein incorporated by reference. Fostel et al. disclose antifungalcompounds including Corynecandin, Mer-WF3010, Fusacandins,Artrichitin/LL 15G256γ, Sordarins, Cispentacin, Azoxybacillin,Aureobasidin and Khafrefungin.

Lipopeptide compounds may be administered according to this method untilthe bacterial infection is eradicated or reduced. In one embodiment, alipopeptide compound is administered for a period of time from 3 days to6 months. In a preferred embodiment, a lipopeptide compound isadministered for 7 to 56 days. In a more preferred embodiment, alipopeptide compound is administered for 7 to 28 days. In an even morepreferred embodiment, a lipopeptide compound is administered for 7 to 14days. Lipopeptide compounds may be administered for a longer or shortertime period if it is so desired.

General Procedures for Lipopeptide Compound Synthesis

Lipopeptide compounds of Formula I may be produced as described below.The lipopeptide compounds of the instant invention may be producedsemi-synthetically using daptomycin as a starting point or may beproduced by a total synthesis approach.

For the semi-synthetic approach according to the present invention,daptomycin may be prepared by any method known in the art. See, e.g.,U.S. Pat. Nos. 4,885,243 and 4,874,843. Daptomycin may be used in itsacylated state or it may be deacylated prior to its use as describedherein. Daptomycin may be deacylated using Actinoplanes utahensis asdescribed in U.S. Pat. No. 4,482,487. Alternatively, daptomycin may bedeacylated as follows:

Daptomycin (5.0 g) was dissolved in water (25 ml) and adjusted to pH 9with 5M sodium hydroxide. Ditert-butyldicarbonate (1.5 g) was added andthe mixture was adjusted to maintain pH 9 with 5 M sodium hydroxideuntil the reaction was complete (4 hours). The pH was adjusted to 7 andthe mixture was loaded onto a Bondesil 40μ C8 resin column. The columnwas washed with water and the product was eluted from the column withmethanol. Evaporation of the methanol gave BOC-protected daptomycin as ayellow powder.

A preparation of deacylase enzyme was produced from recombinantStreptomyces lividans, which expresses the Actinoplanes utahensisdeacylase enzyme. The enzyme in ethylene glycol (400 μ1) was added toBOC-protected daptomycin (1 g) in water (100 ml) at pH 7-8. Afterincubation for 72 hours, the mixture was loaded on a Bondesil 40μ C8resin column. The column was washed with water and the product waseluted from the column with 10% acetonitrile in water. The product wasevaporated to give deacylated BOC-protected daptomycin as a yellowpowderKynurenine Derivatives

Daptomycin can be converted into analogs bearing modifications at the R²position by converting the aromatic amino group to the diazonium saltcompound I with reagents such as sodium nitrite/hydrochloric acid orisoamylnitrite. Using chemistry known to those skilled in the art andfollowing the teachings of the disclosure, the diazonium group can thenbe displaced by reagents such as sodium azide, potassium ethylxanthateor copper chloride to yield derivative compounds II, wherein R¹⁹ is aspreviously defined.

Additionally, compound I can be converted to the azide compound III byreaction with an azide source, typically sodium azide. Modifications tothe ketone group can then be undertaken using chemistry known to thosehaving ordinary skill in the art, such as reduction, oxime formation,ketalization conversion to a leaving group and displacement to givecompounds of formula IV, wherein R¹⁷ and R¹⁸ are as previously defined.

Compound IV may also be converted to compound V by reducing the azidegroup to the amine using chemistry known to those having ordinary skillin the art, and following the teachings of the disclosure, such asreaction with triphenyl phosphine and water, or reducing agents such assodium borohydride wherein R¹⁷ and R¹⁸ are as previously defined.

Additionally compound I can be converted into compound VI by reductionwith hypophosphorus acid. Modifications to the ketone group can then beundertaken using chemistry known to those having ordinary skill in theart similar to those used in scheme 2, wherein R¹⁷ and R¹⁸ are aspreviously defined.Ornithine Derivatives

Daptomycin can be converted into analogs bearing modifications at the R¹position by treating the amino group of the ornithine with reagents suchas isocyanates, isothiocyanates, activated esters, acid chlorides,sulfonylchlorides or activated sulfonamides, heterocycles bearingreadily displaceable groups, imidates, lactones or reductively withaldehydes to yield compound VIII, wherein R¹ is as previously defined.Tryptophan Amine Derivatives

Daptomycin can be converted into compound IX by first protecting theornithine amine with an appropriate amino protecting group (P) known tothose skilled in the art and following the teachings of the disclosure.The decyl side chain on the tryptophan is then removed using an enzymecapable of deacylating daptomycin, such as that described above.

Compound IX can be modified at the tryptophan amine with reagents suchas isocyanates, isothiocyanates, activated esters, acid chlorides,sulfonylchlorides or activated sulfonamides, heterocycles bearingreadily displaceable groups, imidates, lactones or reductively withaldehydes to yield compound X. Compound X can be deprotected to givecompound XI according to procedures known to those skilled in the artfollowing the disclosure of this invention, wherein R is as previouslydefined.

The above modifications to the ornithine amine R¹, tryptophan amine R orkynurenine side chain R² may be independently combined to yieldadditional compounds that are modified at up to all three sites. Inorder to achieve these modifications, it may be necessary to protectcertain functionalities in the molecule. Protecting thesefunctionalities should be within the expertise of one skilled in the artfollowing the disclosure of this invention. See, e.g., Greene, supra.

Solid Support Synthesis of Lipopeptide Compounds

In an alternative embodiment of the invention, the lipopeptide compoundsof Formula I may be synthesized on a solid support as outlined below. Instep 1, a suitably N-protected-βMeGlu(OH)—OAllyl ester is coupled to asuitable resin to give Compound XII. Deprotection of the amino group ofCompound XII, followed by coupling of the amino group with a suitablyprotected seryl derivative (Al) gives Compound XIII, wherein P is asuitable protecting group. This peptide coupling process, i.e.,deprotection of the alpha-amino group, followed by coupling to asuitably protected amino acid, is repeated until the desired number ofamino acids have been coupled to the resin. In the scheme shown below,eleven amino acids have been coupled to give Compound XIV. Addition ofan activated R group, R*, is added to Compound XIV to give Compound XV.In step 4, Compound XV is cyclized to give Compound XVI. Subsequently,in step 5, Compound XVI is removed from the resin to give thelipopeptide Compound XVII.

wherein A¹, is a suitably protected serine derivative, wherein R³¹ is asuitable, cleavable hydroxyl protecting group as outlined below.

wherein A² and A⁷, are suitably protected glycine derivatives asoutlined below.

wherein A³ , A⁵ and A⁹ are suitably protected aspartic acid derivativesas outlined below, wherein ²⁸R, ²⁹R and ³⁰R are cleavable protectinggroups, preferably t-butyl groups.

wherein A⁴ is a suitably protected alanine derivative as outlined below.

wherein A⁶ is a suitably protected ornithine derivative as outlinedbelow, or derivatized ornthine wherein *R¹ is R¹ as previously describedor alternatively a protected form of R¹ that would yield R¹ uponsubsequent deprotection.

wherein A⁸ is a suitably protected depsipeptide as outlined below, Y isa protecting group that is cleavable under conditions that leave otherprotecting groups intact to the others used, i.e., Alloc; and wherein*R² is R² as previously described or alternatively a protected form ofR² that would yield R² upon subsequent deprotection. Preferably ²*R is akynurenine, or substituted kynurenine side chain, most preferably

wherein A¹⁰ is a suitably protected asparagine derivative as outlinedbelow.

wherein A¹¹ is a suitably protected tryptophan derivative as koutlinedbelow, wherein R*³⁷ is hydrido or a suitable protecting group,preferably t-butoxy carbonyl.

It will be understood by those skilled in the art that both the aminoand the side chain functional groups must be suitably protected prior toattaching them to the growing peptide chain. Suitable protecting groupscan be any group known in the art to be useful in peptide synthesis.Such pairings of protecting groups are well known See, e.g., “SynthesisNotes” in the Novabiochem Catalog and Peptide Synthesis Handbook (1999),pages S1-S93 and references cited therein. Following the disclosure ofthe present application, the selection of protecting groups and methodof use thereof will be known to one skilled in the art.

It will also be understood by those skilled in the art that the choiceof protecting group on the side chain functional groups will eitherresult or not result in the protecting group being cleaved concomitantlywith the peptide's final cleavage from the resin, which will give thenatural amino acid functionality or a protected derivative thereof,respectively.

The following general procedures serve to exemplify the solid supportsynthesis of compounds of Formula I.

Step 1: Coupling Suitably-N-protected,β-MeGlu(OH)—OAllyl Ester to aResin

Five molar equivalents each, with respect to the resin, of asuitably-N-protectedβMeGlu(OH)—OAllyl ester, 1,3-Diisopropylcarbodiimide(DIC) and 1-Hydroxy-7-azabenzotriazole (HOAt) are stirred for 30 mins indimethylformamide (DMF; 5 ml/g resin). A suitably functionalised resinor solid support, such as, but not limited to, Wang, Safety Catch, Rink,Knorr, PAL, or PAM resin, is added and the resulting suspension isstirred for 16 hrs. The resin-N-protected βMeGlu(OH)—OAllyl ester isthen filtered, dried and the coupling is repeated. The N-protectinggroup is then removed using the appropriate conditions given in thecoupling steps below.

Step 2: (A) General Coupling Cycle for Amino Acids with anN-9-Fluorenylmethoxycarbonyl (Fmoc) Protecting Group

Five molar equivalents each, with respect to the resin-AA(whereinresin-AA is defined as the resin attached the the growing amino acidchain), of a suitably protected Fmoc amino acid, DIC, and HOAt (0.5molar solution in DMF) are added to the resin-AA, along with sufficientDMF to give a working volume. The mixture is shaken for one hour,filtered, and the coupling is repeated. After the second coupling theresin is washed twice with DMF, twice with methanol, and twice againwith DMF. The Fmoc group of the newly coupled amino acid A¹⁻¹¹ isdeprotected by stirring the resin product in one working volume of asolution of 20% piperidine in N-methyl pyrolidine for five minutes,filtering the resin, and stirring the resin in 20% piperidine inN-methyl pyrolidine again for 20 minutes. The resin is washed twice withDMF, twice with methanol, and twice again with DMF.

Step 2 (B): General Coupling Cycle of Amino Acids with anN-tert-Butoxycarbonyl (N-Boc) Protecting Group

Five molar equivalents each, with respect to the resin-AA, of a suitablyprotected N-Boc amino acid, DIC, and HOAt (0.5 molar solution in DMF)are added to the resin-AA, along with sufficient DMF to give a workingvolume. The mixture is shaken for one hour, filtered, and the couplingis repeated. After the repeated coupling the resin is washed twice withDMF, twice with methanol, and twice again with DMF. The Boc group of thenewly coupled amino acid A¹⁻¹¹, is then deprotected by stirring theresin in one working volume of CH₂Cl₂:trifluoroacetic acid (TFA) 1:1 for15 minutes, filtering, and stirring in one working volume of CH₂Cl₂:TFA1:1 for another 15 minutes. The resin is neutralized by washing withexcess diisopropylethylamine (DIPEA) in CH₂Cl₂ and then washed twicewith DMF, twice with methanol, and twice again with DMF.

Step 3: Terminal Amine Capping Reaction

Ten molar equivalents, with respect to the resin XV, of a suitablereagent containing R* such as an activated ester, isocyanate,thioisocyanate, anhydride, acid chloride, chloroformate, or reactivesalt thereof, in one working volume of DMF is added to the resin XIV andagitated for 25 hours. The resulting resin XV is washed twice with DMF,twice with methanol, and twice again with DMF.

Step 4: Cyclization

The dried resin XV is placed under an argon atmosphere, and treated witha solution of Pd(PPh₃)₄ 125 mgs/0.1 mmol peptide substrate, in CH₂Cl₂:Acetic acid: N-Methylmorpholine, 40:2:1, 1 ml/0.1 mmol peptidesubstrate. The mixture is stirred for 3 hours at ambient temperature,filtered, and washed twice with DMF, twice with methanol, and twiceagain with DMF. Five molar equivalents each, with respect to the resin,of DIC, and HOAt (0.5 molar solution in DMF) are added to the resin,along with sufficient DMF to give a working volume. The reaction isshaken for 17 hours, filtered, and washed twice with DMF, twice withmethanol, and twice again with DMF to give resin XVI.

Step 5: Cleavage and Isolation of the Lipopeptide

The desired lipopeptide is cleaved from resin XVI and isolated,resulting in a compound in which R²⁷ is OH or NH₂. If Fmoc chemistry isused, the dried resin is suspended in 1 ml/0.1 mmol peptide substrate ofCH₂Cl₂:TFA:Ethanedithiol (EDT):Triusopropylsilane (TIS), 16:22:1:1, andstirred for 6-8 hours at ambient temperature. The resin is filtered,washed with 1 equal volume of cold TFA, and the combined filtrates areevaporated under reduced pressure. Crude product XVII is thenprecipitated by the addition of diethyl ether, and isolated bycentrifugation. This product may be further purified by preparativereverse phase HPLC.

If N-Boc chemistry is used, the dried resin is suspended in hydrogenflouride (HF):anisole:dimethylsulfide (DMS), 10:1:1, and stirred for 2hours at 0° C. The volitiles are evaporated under a stream of nitrogen.The resin is then extracted with TFA, filtered and washed twice withTFA, and the combined TFA filtrates evaporated under reduced pressure.Crude product is then precipitated by the addition of diethyl ether, andisolated by centrifugation. This product may be further purified bypreparative reverse phase HPLC.

If the resin is a Safety Catch resin, then R²⁷═OR or NRH. The driedresin XVI is suspended in N-methylpyrolidine (No) or dimethylsulphoxide(DMSO) (8 ml/g resin), Five equivalents of DIPEA (with respect to resinsubstitution) and 24 equivalents of iodo or bromoacetonitrile (withrespect to resin substitution) are added. The suspension is stirred for24 hours at ambient temperature under inert atmosphere. The resin isfiltered, washed with tetrahydrofuran (THF) and DMSO. For an ester, theresin is then treated with an alcohol, hydroxide or alkoxide (20equivalents with respect to resin substitution) in THF for 20 hours. Theresin is filtered, washed with THF and water, and the combined filtratesare evaporated under reduced pressure. Crude product is precipitated bythe addition of diethyl ether, and isolated by centrifugation. Theproduct may be further purified by preparative reverse phase HPLC. Foramides the resin is then treated with a primary or secondary amine (20equivalents with respect to resin substitution) in THF for 12-40 hours,at a gentle reflux under inert atmosphere. The resin is filtered, washedwith THF and water, and the combined filtrates are evaporated underreduced pressure. Crude product is then precipitated by the addition ofdiethyl ether, and isolated by centrifugation. This product may befurther purified by preparative reverse phase HPLC.

In order that this invention may be more fully understood, the followingexamples are set forth. These examples are for the purpose ofillustration only and are not to be construed as limiting the scope ofthe invention in any way.

EXAMPLE 1 Preparation of Compounds 1, 19, 40-44, 49, 72-74, 100, 115-116and 123-125

Daptomycin (5.0 g) was dissolved in water (25 ml) and adjusted to pH 9with 5M sodium hydroxide. Ditertbutyldicarbonate (1.5 g) was added andthe mixture was adjusted to maintain pH 9 with 5 M sodium hydroxideuntil the reaction was complete (4 hours). The pH was adjusted to 7 andthe mixture was loaded onto a Bondesil 40μ C8 resin column. The columnwas washed with water and the product was eluted from the column withmethanol. Evaporation of the methanol gave BOC-protected daptomycin(5.08 g) as a yellow powder.

A preparation of deacylase enzyme was produced from recombinantStreptomyces lividans, which expresses the Actinoplanes utahensisdeacylase enzyme. The enzyme in ethylene glycol (400 μl) was added toBOC-protected daptomycin (1 g) in water (100 ml) at pH 7-8. Afterincubation for 72 hours, the mixture was loaded on a Bondesil 40μ C8resin column. The column was washed with water and the product waseluted from the column with 10% acetonitrile in water. The product wasevaporated to give deacylated BOC-protected daptomycin (440 mg) as ayellow powder.

Deacylated BOC-protected daptomycin (100 mg) and octyl isocyanate (20μl) were stirred at room temperature in dry dimethylformamide (5 ml) for24 hours. Evaporation of the solvent gave a yellow powder residue, whichwas stirred in a mixture of trifluoroaceticacid/dichloromethane/triisopropylsilane/ethane dithiol (11/8/0.5/0.5) (2ml) for 2 hours. Evaporation gave a yellow residue that was purified bypreparative HPLC on an IBSILC-8 5μ 250×20.2 mm column. The column waseluted at 20 ml/min with 36% acetonitrile in 5 mM ammonium phosphatebuffer. Collected fractions were freeze-dried. The freeze-dried residuewas dissolved in water (5 ml) and applied to a Bondesil 40μ C8 resincolumn. The column was washed with water and eluted with methanol.Evaporation of the methanol gave compound 1 as a pale yellow solid (30mg).

In an analogous manner, compounds 19, 40-44, 49, 72-74, 100, 115-116 and123-125 can be prepared by those having ordinary skill in the artfollowing the teachings of the disclosure as detailed in the aboveexample by appropriate substitutions of reagents.

EXAMPLE 1a Preparation of Compounds 18, 37-39, 45-47

Deacylated BOC-protected daptomycin (100 mg) and 4-chloro-4-biphenylacetic acid pentafluorophenyl ester (32 mg) were stirred in drydimethylformamide (3 ml) at room temperature for two days. The mixturewas loaded on an IBSIL-C8 5μ 250×20.2 mm column and was eluted at 20ml/min with 37% acetonitrile in 5 mM ammonium phosphate buffer.Fractions containing the desired compound were collected andfreeze-dried. The freeze-dried residue was dissolved in water (5 ml) andapplied to a Bondesil 40μ C8 resin column, washed with water and elutedwith methanol. Evaporation of the methanol gave the Boc-protectedintermediate as a pale yellow solid (41 mg).

The Boc-protected intermediate (40 mg) was stirred in trifluoroaceticacid (2 ml) and anisole (0.1 ml) at room temperature for 2 hours.Removal of the solvents under reduced pressure gave a residue which wasloaded onto an IBSIL-C8 5μ 250×20.2 mm column and was eluted at 20ml/min with 37% acetonitrile in 5 mM ammonium phosphate buffer.Fractions containing the desired compound were collected andfreeze-dried. The freeze-dried residue was dissolved in water (5 ml) andapplied to a Bondesil 40μ C8 resin column, washed with water and elutedwith methanol. Evaporation of the methanol gave compound 18 as a paleyellow solid (10 mg).

In an analogous manner, compounds 37-39 and 45-47 can be prepared bythose having ordinary skill in the art following the teachings of thedisclosure as detailed abovein the above example by appropriatesubstitution of reagents.

EXAMPLE 1b Preparation of Compounds 110, 112, 109 and 111

Boc-protected daptomycin α,β-tridecenoyl amide (compound 110) wasprepared from deacylated Boc-protected daptomycin α,β-tridecenoylpentafluorophenol ester according to Examples 1 and 1a. Compound 110(0.21 g) in dry dichloromethane (8 ml), trifluoroacetic acid (11 ml) andethane dithiol (0.25 ml) was stirred for 3 hours at room temperature.Concentration under reduced pressure gave a light brown oil which waspurified on an IBSIL-C8 5μ 250×20.2 mm column and eluted at 25 ml/minwith 30-60% acetonitrile in 5 mM ammonium phosphate gradient over 40minutes. Fractions containing the desired compound were collected andfreeze-dried. The freeze-dried residue was dissolved in water (5 ml) andapplied to a Bondesil 40μ C8 resin column, washed with water and elutedwith methanol. Evaporation of the methanol gave compound 112 (53.8 mg)as a pale yellow solid.

In an analogous manner, compounds 109 and 111 can be prepared by thosehaving ordinary skill in the art following the teachings of thedisclosure as detailed abovein the above example by appropriatesubstitution of reagents.

EXAMPLE 2 Preparation of Compound 2

Dodecyl isocyanate (0.507 g) in dry dimethylformamide (3 ml) was addedto deacylated Boc-protected daptomycin (3.14 g) in dry dimethylformamide(30 ml). The mixture was stirred at room temperature under nitrogen.After 7 hours the mixture was purified on a Bondesil 40μ C8 resin columnwith 10% acetonitrilewater followed by 50% acetonitrilewater. Thedesired fractions were freeze-dried to give Boc-protected daptomycindodecyl urea (3.38 g) as pale yellow fluffy solid.

Boc-protected daptomycin dodecyl urea (2.42 g) in dry dichloromethane(20 ml), trifluoroacetic acid (22 ml) and ethane dithiol (0.5 ml) wasstirred for 4 hours at room temperature. The mixture was concentrated toa light brown oil then triturated with methanol and diethyl ether. Afterthe mixture was centrifuged the yellow residue was loaded on an IBSIL-C85μ 250×20.2 mm column and eluted at 25 ml/min with 30-60% acetonitrilein 5 mM ammonium phosphate gradient over 40 minutes. Fractionscontaining the desired compound were collected and freeze-dried. Thefreeze-dried residue was dissolved in water (5 ml) and applied to aBondesil 40μ C8 resin column, washed with water and eluted withmethanol. Evaporation of the methanol gave compound 2 (2.53 g) as paleyellow solid.

EXAMPLE 2a Preparation of Compound 48

Undecyl isocyanate (0.197 g) in dry dimethylformamide (1 ml) was addedto deacylated Boc-protected daptomycin (1.62 g) in dry dimethylformamide(20 ml). The mixture was stirred at room temperature under nitrogen for7 hours. The mixture was then purified on a Bondesil 40μ C8 resin columnwith 10% acetonitrile water followed by 50% acetonitrilewater. Thedesired fractions were freeze-dried to give Boc-protected daptomycinundecyl urea (1.58 g) as pale yellow fluffy solid.

Boc-protected daptomycin undecyl urea (1.58 g) in dry dichloromethane(20 ml), trifluoroacetic acid (22 ml) and 5% anisole was stirred for 4hours before being evaporated to dryness. The residue was loaded on anIBSIL-C8 5μ 250×20.2 mm column and eluted at 25 ml/min with 30-60%acetonitrile in 5 mM ammonium phosphate gradient over 40 minutes.Fractions containing the desired compound were collected andfreeze-dried. The freeze-dried residue was dissolved in water (5 ml) andapplied to a Bondesil 40μ C8 resin column, washed with water and elutedwith methanol. Evaporation of the methanol gave compound 48 (136.5 mg)as pale yellow solid.

EXAMPLE 2b Preparation of compounds 117 and 118

Nonyl isocyanate (40.6 mg) in dry dimethylformamide (0.2 ml) was addedto deacylated Boc-protected daptomycin (313.2 mg) in drydimethylformamide (2ml) The mixture was stirred at room temperatureunder nitrogen. After 7 hours the mixture was purified on an IBSIL-C8 5μ250×20.2 mm column and eluted at 25 ml/min with 30-60% acetonitrile in 5mM ammonium phosphate gradient over 40 minutes. Fractions containing thedesired compound were collected at 21 minutes and freeze-dried. Thefreeze-dried residue was dissolved in water (5 ml) and applied to aBondesil 40μ C8 resin column, washed with water and eluted withmethanol. Evaporation of the methanol gave compound 117 (158.8 mg) aspale yellow solid.

Compound 117 (58.9 mg) in dry dichloromethane (5 ml), trifluoroaceticacid (2 ml) and ethane dithiol (0.05 ml). The mixture was stirred for 2hours at room temperature before being evaporated to dryness. Theresidue was loaded on an IBSIL-C8 5μ 250×20.2 mm column and eluted at 25ml/min with 30-60% acetonitrile in 5 mM ammonium phosphate gradient over40 minutes. Fractions containing the desired compound were collected andfreeze-dried. The freeze-dried residue was dissolved in water (5 ml) andapplied to a Bondesil 40μ C8 resin column, washed with water and elutedwith methanol. Evaporation of the methanol gave compound 118 (11.2 mg)as pale yellow solid.

EXAMPLE 2c Preparation of Compounds 119 and 120

Decyl isocyanate (0.44 g) in dry dimethylformamide (0.2 ml) was added todeacylated Boc-protected daptomycin (3.13 g) in dry dimethylformamide(20 ml). The mixture was stirred at room temperature under nitrogen.After 7 hours the mixture was purified on a Bondesil 40μ C8 resin columnwith 10% acetonitrile-water followed by 50% acetonitrilewater. Thedesired fractions were freeze-dried to give compound 119 (1.73 g) aspale yellow solid.

Compound 119 (1.73 g) in dry dichloromethane (20 ml), trifluoroaceticacid (22 ml) and ethane dithiol (0.5 ml) was stirred for 4 hours at roomtemperature before being evaporated to dryness. The residue wastriturated with methanol and diethyl ether then loaded on an IBSIL-C8 5μ250×20.2 mm column and eluted at 25 ml/min with 30-60% acetonitrile in 5mM ammonium phosphate gradient over 40 minutes Fractions containing thedesired compound were collected and freeze-dried. The freeze-driedresidue was dissolved in water (5 ml) and applied to a Bondesil 40 A C8resin column, washed with water and eluted with methanol. Evaporation ofthe methanol gave compound 120 (359.8 mg) as pale yellow solid.

EXAMPLE 3 Preparation of Compounds 3, 5-6, 8-13, 20-24, 34-36, 50, 71and 75

Daptomycin (250 mg) and N-tBoc-L-tryptophan-p-nitrophenyl ester (144 mg)were stirred in dry dimethylformamide (3 ml) at room temperature for twodays. The mixture was loaded on an IBSIL-C8 5μ 250×20.2 mm column andwas eluted at 20 ml/min with 37% acetonitrile in 5 mM ammonium phosphatebuffer. Fractions containing the desired compound were collected andfreeze-dried. The freeze-dried residue was dissolved in water (5 ml) andapplied to a Bondesil 40μ C8 resin column, washed with water and elutedwith methanol. Evaporation of the methanol gave N-Boc tryptophandaptomycin as a pale yellow solid (130 mg).

A preparation of deacylase enzyme was produced from recombinantStreptomyces lividans, which expresses the Actinoplanes utahensisdeacylase enzyme. The enzyme in ethylene glycol (400 μl) was added tothe solution of N-Boc tryptophan daptomycin (100 mg) in HPLC grade water(20 ml). The solution was adjusted to pH 8.5 with sodium hydroxide (1M). The mixture was stirred for 24 hours. The mixture was loaded on a C8resin plug column, washed with water and eluted with methanol.Evaporation of the methanol gave a residue which was applied to anIBSIL-C8 5μ 250×20.2 mm column and was eluted at 20 ml/min with 37%acetonitrile in 5 mM ammonium phosphate buffer. Fractions containing thedesired compound were collected and freeze-dried. The freeze-driedresidue was dissolved in water (5 ml) and applied to a Bondesil 40μ C8resin column, washed with water and eluted with methanol. Evaporation ofthe methanol gave deacylated N-Boc tryptophan daptomycin as a paleyellow solid (42 mg).

Deacylated N-Boc tryptophan daptomycin (20 mg) was stirred in drydimethylformamide (2 ml) at room temperature. Undecyl isocyanate (2.25mg) was added to the solution. After stirring at ambient temperature for24 hours, the mixture was diluted with water (10 ml) and applied to aBondesil 40μ C8 resin column, washed with water and eluted withmethanol, Evaporation of the methanol gave the undecyl urea of N-Boctryptophan daptomycin as a pale yellow solid (21 mg).

N-Boc tryptophan daptomycin undecyl urea (21 mg) was stirred intrifluoroacetic acid (2 ml) and anisole (0.1 ml) at room temperature for2 hours. Removal of the solvents under reduced pressure gave a residuewhich was loaded on an IBSIL-C8 5μ 250×20.2 mm column and eluted at 20ml/min with 37% acetonitrile in 5 mM ammonium phosphate buffer.Fractions containing the desired compound were collected andfreeze-dried. The freeze-dried residue was dissolved in water (5 ml) andapplied to a Bondesil 40μ C8 resin column, washed with water and elutedwith methanol. Evaporation of the methanol gave compound 3 as a paleyellow solid (0.8 mg).

In an analogous manner, compounds 5-6, 8-13, 20-24, 34-36, 50, 71 and 75can be prepared as detailed in the above example by appropriatesubstitutions of reagents.

EXAMPLE 3a Preparation of Compound 7

Deacylated N-Boc tryptophan daptomycin (50 mg) and nonaldehyde (4.1 mg)were stirred in dry dimethylformamide (2 ml) at room temperature. Sodiumtriacetoxy borohydride (3.6 mg) was added to the solution. The mixturewas stirred for 24 hours, then loaded on an IBSIL-C8 5μ 250×20.2 mmcolumn and eluted at 20 ml/min with 37% acetonitrile in 5 mM ammoniumphosphate buffer. Fractions containing the desired compound werecollected and freeze-dried. The freeze-dried residue was dissolved inwater (5 ml) and applied to a Bondesil 40μ C8 resin column, washed withwater and eluted with methanol. Evaporation of the methanol gave nonylamino N-Boc tryptophan daptomycin as a pale yellow solid (14 mg).

Nonyl amino N-Boc tryptophan daptomycin (14 mg) was stirred intrifluoroacetic acid (2 ml) and anisole (0.1 ml) at room temperature for2 hours. Removal of the solvents under reduced pressure gave a residuewhich was loaded on an IBSIL-C8 5μ 250×20.2 mm column and was eluted at20 ml/min with 37% acetonitrile in 5 mM ammonium phosphate buffer.Fractions containing the desired compound were collected andfreeze-dried. The freeze-dried residue was dissolved in water (5 ml) andapplied to a Bondesil 40μ C8 resin column, washed with water and elutedwith methanol. Evaporation of the methanol gave compound 7 as a paleyellow solid (5 mg)

EXAMPLE 3b Preparation of Compound 17

Deacylated N-Boc tryptophan daptomycin (50 mg) was stirred in drydimethylformamide (2 ml) at room temperature. Dodecyl isocyanate (6.0mg) was added to the solution. The mixture was stirred for 24 hours. Themixture was loaded on an IBSIL-C8 5μ 250×20.2 mm column and eluted at 20ml/min with 37% acetonitrile in 5 mM ammonium phosphate buffer.Fractions containing the desired compound were collected andfreeze-dried. The freeze-dried residue was dissolved in water (5 ml) andapplied to a Bondesil 40μ C8 resin column, washed with water and elutedwith methanol. Evaporation of the methanol gave the dodecyl urea ofN-Boc tryptophan daptomycin as a pale yellow solid (27 mg).

N-Boc tryptophan daptomycin dodecyl urea (25 mg) was stirred intrifluoroacetic acid (2 ml) and anisole (0.1 ml) at room temperature for2 hours. Removal of the solvents under reduced pressure gave a residuewhich was loaded on an IBSIL-C8 5μ 250×20.2 mm column and eluted at 20ml/min with 37% acetonitrile in 5 mM ammonium phosphate buffer.Fractions containing the desired compound were collected andfreeze-dried. The freeze-dried residue was dissolved in water (5 ml) andapplied to a Bondesil 40μ C8 resin column, washed with water and elutedwith methanol. Evaporation of the methanol gave compound 17 as a paleyellow solid (4.3 mg).

EXAMPLE 4 Preparation of Compounds 69, 25, 56-58, 62-64, 70, 106 and 108

Daptomycin octyl urea synthesized from deacylated Boc-protecteddaptomycin by using octyl isocyanate according to examples 1 and 1a (60mg) and N-tBoc-L-tryptophan-p-nitrophenyl ester (31 mg) were stirred indry dimethylformamide (2 ml) at room temperature for two days. Themixture was loaded onto an IBSIL-C8 5μ 250×20.2 mm column and was elutedat 20 ml/min with 37% acetonitrile in 5 mM ammonium phosphate buffer.Fractions containing the desired compound were collected andfreeze-dried. The freeze-dried residue was dissolved in water (5 ml) andapplied to a Bondesil 40μ C8 resin column, washed with water and elutedwith methanol. Evaporation of the methanol gave the acylatedintermediate as a pale yellow solid (29 mg).

The acylated intermediate (25 mg) was stirred in trifluoroacetic acid (2ml) and anisole (0.1 ml) at room temperature for 2 hours. Evaporationunder reduced pressure gave a residue which was loaded on an IBSIL-C8 5μ250×20.2 mm column and was eluted at 20 ml/min with 37% acetonitrile in5 mM ammonium phosphate buffer. Fractions containing the desiredcompound were collected and freeze-dried. The freeze-dried residue wasdissolved in water (5 ml) and applied to a Bondesil 40μ C8 resin column,washed with water and eluted with methanol. Evaporation of the methanolgave compound 69 as a pale yellow solid (5 mg).

In an analogous manner, compounds 25, 56-58, 62-64, 70, 106 and 108 canbe prepared by those having skill in the art as detailed in the aboveexample by appropriate substitutions of reagents.

EXAMPLE 4a Preparation of Compounds 89, 76-78, 87-88 and 113

Daptomycin dodecyl urea synthesized from deacylated Boc-protecteddaptomycin by using dodecyl isocyanate according to examples 1 and 1a(200 mg) and 2-imidazolecarboxaldehyde (21 mg) in dry dimethylformamide(1.0 ml) was added sodium triacetoxyborohydride (152 mg). The mixturewas stirred at room temperature for 24 hours before purification bypreparative HPLC. The mixture was loaded on an IBSIL-C8 5μ 250×20.2 mmcolumn and eluted at 25 ml/min with 30-60% acetonitrile in 5 mM ammoniumphosphate gradient over 30 minutes. The desired fractions were collectedat 21 minutes and freeze-dried. The freeze-dried residue was dissolvedin water (3 ml) and applied to a plug of Bondesil 40μ C8 resin (500 mg).The Bondesil resin was washed with water (10 ml) and then the productwas eluted with methanol (10 ml). Evaporation of the methanol gavecompound 89 as a pale yellow solid (15 mg).

In an analogous manner, compounds 76-78, 87-88 and 113 can be preparedby those having ordinary skill in the art by following the teachings ofthe disclosure as detailed in the above example by appropriatesubstitutions of reagents.

EXAMPLE 4b Preparation of Compound 114

Daptomycin undecyl urea synthesized from deacylated Boc-protecteddaptomycin using undecyl isocyanate according to examples 1 and 1a (100mg), and 5-methoxyindole-3-carboxaldehyde (11 mg) in drydimethylformamide (0.6 ml) was added sodium triacetoxyborohydride (76mg). The mixture was stirred at room temperature for 24 hours beforepurification by preparative HPLC. The mixture was loaded on an IBSIL-C85μ 250×20.2 mm column and eluted at 25 ml/min with 30-60% acetonitrilein 5 mM ammonium phosphate gradient over 30 minutes. The desiredfractions were collected at 21 minutes and freeze-dried. Thefreeze-dried residue was dissolved in water (2 ml) and applied to a plugof Bondesil 40μ C8 resin (500 mg). The Bondesil resin was washed withwater (10 ml) and then the product was eluted with methanol (10 ml).Evaporation of the methanol gave compound 114 as a pale yellow solid (10mg).

EXAMPLE 5

Compounds according to Formula I were tested for antimicrobial activityagainst a panel of organisms according to standard procedures describedby the National Committee for Clinical Laboratory Standards (NCCLSdocument M7-A5, Vol. 20, No. 2, 2000) except that all testing wasperformed at 37° C. Compounds were dissolved in 100% dimethyl sulfoxideand were diluted to the final reaction concentration (0.1 μg/mL-100μg/mL) in microbial growth media. In all cases the final concentrationof dimethyl sulfoxide incubated with cells is less than or equal to 1%.For minimum inhibitory concentration (MIC) calculations, 2-folddilutions of compounds were added to wells of a microtiter platecontaining 5×10⁴ bacteria cells in a final volume of 100 μL of media(Mueller-Hinton Broth supplemented with 50 mg/L Ca²⁺). The opticaldensities (OD) of the bacterial cells, which measures bacterial cellgrowth and proliferation, were measured using a commercial plate reader.The MIC value is defined as the lowest compound concentration inhibitinggrowth of the test organism. The MIC (in μg/ml) values of representativecompounds of the present invention are listed in Table VI.

EXAMPLE 6

The in vivo antibacterial activity of Compound 2 (see Table IV) wasestablished by infecting female CD-1 mice (Charles River Lab, Mass. )weighing 19-23 g intraperitoneally with Methicillin Resistant S. aureus(MRSA) inoculum. The inoculum was prepared from Methicillin Resistant S.aureus (ATCC 43300). The MRSA inoculum was cultured in Mueller-Hinton(MH) broth at 37° C. for 18 hours. The optical density at 600 nm (OD₆₀₀)was determined for a 1:10 dilution of the overnight culture. Bacteria(8×10⁸ cfu) was added to 20 ml of phosphate buffered saline (SigmaP-0261) containing 6% hog gastric mucin (Sigma M-2378). Group 1-5animals were injected with 0.5 ml of the inoculum, equivalent to 2×10⁷cfu/mouse, which is the dose causing ˜100% death of the animals withouttreatment. Group 6 animals received no inoculum.

The test compound (10 mg) was dissolved in 10.0 ml of 50 mM phosphatebuffer to give a solution of 1 mg/ml (pH=7.0). This solution wasserially diluted with vehicle by 4-fold (1.5 ml to 6 ml) to give 0.25,0.063 and 0.016 mg/ml solutions. All the solutions were filtered with0.2 μm Nalgene syringe filter. Immediately after the bacterialinoculation, group 1 animals were subcutaneously (sc) injected withbuffer (no test compound) and groups 2 to 5 were given test compound scat 10, 2.5, 0.63, and 0.16 mg/kg, respectively. Group 6 animals compound2 s.c at 10 mg/kg only. These injections were repeated once at 4 hoursafter the inoculation for the respective groups. The injection volume ateach time was 10 ml per kilogram of body weight. The results of the invivo efficacy test are summarized in Table IV, which provides arepresentative example of the results obtained for Compound 2. The 50%effective dose (ED₅₀) is calculated on the basis of the number of micesurviving 7 days after inoculation. The ED₅₀ in mg/kg of otherrepresentative compounds of the present invention are listed in Table V.

TABLE V Survival Group # of mice Inoculated with Treatment (7 days) 1 5MRSA #43300 Phosphate buffer 0/5 2 × 10⁷ cfu/mouse 10 ml/kg, s.c. x2 2 5MRSA #43300 Compound 2 5/5 2 × 10⁷ cfu/mouse 10 mg/kg, s.c. x2 3 5 MRSA#43300 Compound 2 5/5 2 × 10⁷ cfu/mouse 2.5 mg/kg, s.c. x2 4 5 MRSA#43300 Compound 2 5/5 2 × 10⁷ cfu/mouse 0.63 mg/kg, s.c. x2 5 5 MRSA#43300 Compound 2 1/5 2 × 10⁷ cfu/mouse 0.16 mg/kg, s.c. x2 6 5 NOCompound 2 5/5 10 mg/kg s.c. x2The ED₅₀ of compound 2 is calculated to be 0.43 mg/kg.

The ED₅₀ was determined for other compounds of this invention in asimilar manner.

TABLE VI MIC MIC Compound (μg/ml) (μg/ml) ED₅₀₎ # S. aureus E. faecalis(mg/kg) 1 ++ + 2 +++ ++ +++ 3 +++ +++ +++ 5 ++ ++ 6 + 7 + + 8 ++ + 9++ + 10 ++ + 11 + + 12 + + 13 ++ + 17 +++ +++ 18 +++ ++ +++ 19 20 + 21 +22 + 23 ++ 24 +++ + 25 ++ + 34 +++ ++ 35 ++ + 36 + 37 ++ + 38 +++ ++ 39+++ +++ 40 + 41 43 44 ++ + 45 +++ ++ 46 ++ + 47 +++ +++ 48 +++ +++ +++49 ++ ++ 50 +++ ++ 56 ++ + 57 ++ ++ 58 ++ ++ 62 ++ + 63 +++ ++ 64 +++ ++69 +++ + 70 71 +++ + 72 +++ + 73 +++ ++ 74 +++ ++ 75 +++ ++ 76 ++ + 77+++ + 78 ++ + 87 +++ ++ 88 +++ ++ 89 +++ +++ 100 + 106 ++ + 108 ++ 109++ + 110 +++ ++ 111 +++ ++ 112 +++ +++ 113 ++ ++ 114 +++ +++ 115 ++ +116 +++ ++ 117 ++ + 118 +++ +++ 119 +++ ++ 120 +++ +++ 123 ++ 124 ++ +++125 ++ +++

Wherein “+++” indicates that the compound has an MIC (μg/ml) of 1 μg/mlor less or an ED₅₀ of 1 mg/kg or less;

“++” indicates that the compound has an NEC (μg/ml) or an ED₅₀ (mg/kg)or more than 1 μg/ml or 1 mg/kg, respectively, but less than or equal to10 μg/ml or 10 mg/kg, respectively;

“+” that the compound has an MIC (μg/ml) of greater than 10 μg/ml or anED₅₀ of greater than 10 mg/kg; and

wherein a blank indicates that the MIC or ED₅₀ was not determined.

All publications and patent applications cited in this specification areherein incorporated by reference as if each individual publication orpatent application were specifically and individually indicated to beincorporated by reference. Although the foregoing invention has beendescribed in some detail by way of illustration and example for purposesof clarity of understanding, it will be readily apparent to those orordinary skill in the art in light of the teachings of this inventionthat certain changes and modifications may be made thereto withoutdeparting from the spirit or scope of the appended claims.

1. A compound having the formula (I):

wherein X and X″ are independently selected from C═O, C═S, C═NH,C═NR^(X), S═O or SO₂; wherein R^(X) is selected from alkyl, alkenyl,alkynyl, aryl, heteroaryl, cycloalkyl, heterocyclyl, hydroxyl, alkoxy,carboxy or carboalkoxy; wherein B is X″R^(Y), H, alkyl, alkenyl,alkynyl, aryl, heteroaryl, cycloalkyl and heterocyclyl; and whereinR^(Y) is selected from hydrido, alkyl, alkenyl, alkynyl, aryl,heteroaryl, cycloalkyl, heterocyclyl or hydroxyl; wherein A is H, NH₂,NHR^(A), NR^(A)R^(B), heteroaryl, cycloalkyl or heterocyclyl; whereinR^(A) and R^(B) are independently selected from alkyl, alkenyl, alkynyl,aryl, heteroaryl, cycloalkyl, heterocyclyl or carboalkoxy; provided thatwhen B is H and X is C═O, then A is other than (a) a pyridinyl ringsubstituted with a single NHC(O)R^(D) substitutent or (b) a (C₅-C₆)saturated cycloalkyl ring substituted with a single NHC(O)R^(D)substitutent, wherein R^(D) is (C₂-C₁₇) unsubstituted alkyl or (C₂-C₁₇)unsubstituted alkenyl; wherein R¹ is

wherein X′ and X″ are independently selected from C═O, C═S, C═NH,C═NR^(X′), S═O or SO₂; wherein m is 0 or 1; wherein R^(X′) is selectedfrom alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl,heterocyclyl, hydroxyl, alkoxy, carboxy or carboalkoxy; wherein B′ isX′″R^(Y′), H, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl orheterocyclyl; wherein R^(Y′) is selected from hydrido, alkyl, alkenyl,alkylnyl, aryl, heteroaryl, cycloalkyl, heterocyclyl or hydroxyl;wherein A′ is H, NH₂, NHR^(A′), NR^(A′)R^(B)′, alkyl, alkenyl, alkynyl,alkoxy, aryloxy, aryl, heteroaryl, cycloalkyl or heterocyclyl; whereinR^(A′) and R^(B′) are independently selected from alkyl, alkenyl,alkynyl, aryl, heteroaryl, cycloalkyl, heterocyclyl or carboalkoxy;wherein when m is 0, then A′ is additionally selected from the groupconsisting of:

wherein each of R⁵⁰—R⁵³ is independently selected from C₁-C₁₅ is alkyl;alternatively, wherein B′ and A′ together form a 5-7 memberedheterocyclic or heteroaryl ring; wherein R² is

wherein K and K′ together form a C₃-C₇ cycloalkyl or heterocyclyl ringor a C₅-C₁₀ aryl or heteroaryl ring; wherein J is selected from thegroup consisting of hydrido, amino, NHR^(J), NR^(J)R^(K), alkyl,alkenyl, alkynyl, alkoxy, aryloxy, aryl, heteroaryl, cycloalkyl,heterocyclyl, alkylamino, hydroxyl, thio, alkylthio, alkenylthio,sulfinyl, sulfonyl, azido, cyano, halo,

wherein each of R²⁴, R²⁵, and R²⁶ is independently selected from thegroup consisting of alkyl, cycloalkyl, heterocyclyl, aryl andheteroaryl; or R²⁴ and R²⁵ together form a 5-8 membered heterocyclylring; wherein R^(J) and R^(K) are independently selected from alkyl,alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl or heterocyclyl; oralternatively, wherein J, together with R¹⁷, forms a 5-8 memberedheterocyclyl or cycloalkyl ring; or alternatively, wherein J, togetherwith both R¹⁷ and R¹⁸, forms a 5-8 membered aryl, cycloalkyl,heterocyclyl or heteroaryl ring; and wherein each of R¹⁷ and R¹⁸ isindependently selected from the group consisting of hydrido, halo,hydroxyl, alkoxy, amino, thio, sulfinyl, sulfonyl and

 or wherein R¹⁷ and R¹⁸ taken together can form a group consisting ofketal, thioketal,

wherein each of R²² and R²³ is independently selected from the groupconsisting of hydrido and alkyl.
 2. A compound having the formula (I):

wherein X and X″ are independently selected from C═O, C═S, C═NH,C═NR^(X), S═O or SO₂; wherein R^(X) is selected from alkyl, alkenyl,alkynyl, aryl, heteroaryl, cycloalkyl, heterocyclyl, hydroxyl, alkoxy,carboxy or carboalkoxy; wherein B is X″R^(Y), H, alkyl, alkenyl,alkynyl, aryl, heteroaryl, cycloalkyl or heterocyclyl; and wherein R^(Y)is selected from hydrido, alkyl, alkenyl, alkynyl, aryl, heteroaryl,cycloalkyl, heterocyclyl or hydroxyl; wherein A is aryl; provided thatwhen B is H and X is C═O, then A is other than a phenyl ring substitutedwith either: (a) —O—((C₈-C₁₅) unsubstituted alkyl), wherein said phenylring may be further optionally substituted with one substituent selectedfrom halo, nitro, (C₁-C₃) alkyl, hydroxyl, (C₁-C₃) alkoxy or (C₁-C₃)alkylthio; or (b) —NHC(O)R^(D), wherein the phenyl ring maybe furtheroptionally substituted with 1-2 substituents independently selected fromamino, nitro, (C₁-C₃) alkyl, hydroxyl, (C₁-C₃) alkoxy, halo, mercapto,(C₁-C₃) alkylthio, carbamyl or (C₁-C₃) alkylcarbamyl, wherein R^(D) is(C₁-C₁₇) unsubstituted alkyl or (C₂-C₁₇) unsubstituted alkenyl; whereinR¹ is

wherein X′ and X′″ are independently selected from C═O, C═S, C═NH,C═NR^(X′), S═O or SO₂; wherein m is 0 or 1; wherein R^(X′) is selectedfrom alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl,heterocyclyl, hydroxyl, alkoxy, carboxy or carboalkoxy; wherein B′ isX′″R^(Y′), H, alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl orheterocyclyl; wherein R^(Y′) selected from hydride, alkyl, alkenyl,alkynyl, aryl, heteroaryl, cycloalkyl, heterocyclyl or hydroxyl; whereinA′ is H, NH₂, NHR^(A′), NR^(A′)R^(B′), alkyl, alkenyl, alkynyl, alkoxy,aryloxy, aryl, heteroaryl, cycloalkyl or heterocyclyl; wherein R^(A′)and R^(B′) are independently selected from alkyl, alkenyl, alkynyl,aryl, heteroaryl, cycloalkyl, heterocyclyl or carboalkoxy; wherein whenm is 0, then A′ is additionally selected from the group consisting of:

wherein each of R⁵⁰-R⁵³ is independently selected from C₁-C₁₅ alkyl;alternatively, wherein B′ and A′ together form a 5-7 memberedheterocyclic or heteroaryl ring; wherein R² is

wherein K and K′ together form a C₃-C₇ cycloalkyl or heterocyclyl ringor a C₅-C₁₀ aryl or heteroaryl ring; wherein J is selected from thegroup consisting of hydrido, ammo, NHR^(J), NR^(J)R^(K), alkyl, alkenyl,alkynyl, alkoxy, aryloxy, aryl, heterocyclyl, cycloalkyl, heterocyclyl,alkylamino, hydroxyl, thin, alkylthio, alkenylthio, sulfinyl, sulfonyl,azido, cyano, halo,

wherein each of R²⁴, R²⁵, and R²⁶ is independently selected from thegroup consisting of alkyl, cycloalkyl, heterocyclyl, aryl andheteroaryl; or R²⁴ and R²⁵ together form a 5-8 membered heterocyclylring; wherein R^(J) and R^(K) are independently selected from alkyl,alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl or heterocyclyl; oralternatively, wherein J, together with R¹⁷, forms a 5-8 memberedheterocyclyl or cycloalkyl ring; or alternatively, wherein J, togetherwith both R¹⁷, together with both R¹⁷ and R¹⁸, forms a 5-8 memberedaryl, cycloalkyl, heterocyclyl or heteroaryl ring; and wherein each ofR¹⁷ and R¹⁸ is independently selected from the group consisting ofhydrido, halo, hydroxyl, alkoxy, amino, thio, sulfinyl, sulfonyl and

 or wherein R¹⁷ and R¹⁸ taken together can form a group consisting ofketal, thioketal,

wherein each of R²² and R²³ is independently selected from the groupconsisting of hydrido and alkyl.
 3. The compound according to claim 1,wherein R is selected from the group consisting of:

wherein each of R³, R⁴, R⁵, and R⁶ is independently selected from thegroup consisting of hydrido, alkyl, aryl, heterocyclyl and heteroaryl,and wherein R²⁰⁰ is selected from the group consisting of hydrido,heterocyclyl, and heteroaryl.
 4. The compound according to claim 3,wherein R is selected from

and wherein R^(4′) is selected from the group consisting of heteroaryl,and heterocyclyl.
 5. The compound according to claim 4, wherein R is


6. The compound according to either of claim 1 or 2, wherein R¹ isselected from the group consisting of:

wherein R⁸ is selected from a natural amino acid side chain or an aminoacid side chain; wherein each of R⁹, R¹⁰ and R¹¹ is selected from thegroup consisting of hydrido, alkyl, aryl, heterocyclyl and heteroaryl;wherein R¹² is selected from the group consisting of heterocyclyl,heteroaryl, aryl, and alkyl and wherein R¹³ is selected from(C₁-C₃-alkyl) and aryl.
 7. The compound according to claim 6, wherein R¹is selected from the group consisting of:

wherein R⁸ is selected from tryptophan side chain and lysine side chain;wherein each of R¹⁰ and R¹¹ is independently selected from hydrido andalkyl; wherein R¹² is selected from imidazolyl, N-methylimidazolyl,indolyl, quinolinyl, benzyloxybenzyl, and benzylpiperidenylbenzyl; andwherein X⁴ is fluoro, or trifluoromethyl.
 8. The compound according toeither of claim 1 or 2, wherein J is selected from the group consistingof hydrido, amino, azido and

wherein R¹⁷ and R¹⁸ taken together form a group selected from ketal,

or wherein R¹⁷ is hydroxyl when R¹⁸ is hydrido; or wherein J, togetherwith R¹⁷, forms a heterocyclyl ring.
 9. The compound according to claim8, wherein R² is selected from the group consisting of

 wherein R¹⁷ and R¹⁸ taken together form a group selected from

wherein R²² is selected from the group consisting of H and alkyl; andwherein R¹⁹ is selected from the group consisting of hydrido, amino,azido and


10. The compound according to claim 9, wherein R² is


11. A pharmaceutical composition comprising the compound according toeither of claim 1 or 2 and a pharmaceutically acceptable carrier.
 12. Amethod of treating a bacterial infection in a subject, comprising thestep of administering a therapeutically-effective amount of thepharmaceutical composition according to claim 11 to a subject in needthereof for a time and under conditions effective to ameliorate saidbacterial infection.
 13. The method according to claim 12, wherein saidsubject is selected from the group consisting of a human, an animal, acell culture and a plant.
 14. The method according to claim 12, whereinsaid bacterial infection is caused by a gram-positive bacteria.
 15. Themethod according to claim 14, wherein said bacteria is anantibiotic-resistant bacteria that is resistant to an antibiotic that isnot included within the scope of Formula (I).
 16. The method accordingto claim 15, wherein said antibiotic-resistant bacteria are resistant toan antibiotic selected from the group consisting of vancomycin,methicillin, glycopeptide antibiotics, penicillin and daptomycin. 17.The method according to claim 12, further comprising the step ofco-administering more than one compound of Formula (I) according toeither of claim 1 or 2 to a subject in need thereof.
 18. The methodaccording to claim 12, further comprising the step of co-administering asecond antimicrobial agent wherein said second antimicrobial agent isnot included within the scope of Formula (I).
 19. The method accordingto claim 11, wherein said second antimicrobial agent is selected fromthe group consisting of penicillins, carbapenems, cephalosporins,aminoglycosides, bacitracin, gramicidin, mupirocin, chloramphenicol,thiamphenicol, fusidate sodium, lincomycin, clindamycin, macrolides,novobiocin, polymyxins, rifamycins, spectinomycin, tetracyclines,vancomycin, teicoplanin, streptogramins, anti-folate agents,trimethoprim, pyrimethamine, nitroimidazoles, quinolones,fluoroquinolones, isoniazid, ethambutol, pyrazinamide,para-aminosalicylic acid (PAS), cycloserine, capreomycin, ethionamide,prothionamide, thiacetazone, viomycin, eveminomicin, glycopeptide,glycylcycline, ketolides, oxazolidinone, imipenen, amikacin, netilmicin,fosfomycin, gentamicin, ceftriaxone, ZIRACIN,(56-deacetyl-57-demethyl-45-O-de(2-methyl-1-oxopropyl)-12-O-(2,3,6-trideoxy-3-C-methyl-4-O-methyl-3-nitro-alpha-L-arabino-hexopyranosyl)flambamycin),LY333328 (oritavancin), linezolid(N-[[((5S)-3-[3-fluoro-4-(4-morpholinyl)phenyl]-2-oxo-5-oxazolidinyl]methyl]acetatamide),SYNERCID (dalfopristin-quinupristin), aztreonam(2-[[(Z)-[1-(2-amino-4-thiazolyl)-2-[[(2S,3S)-2-methyl-4-oxo-1-sulfo-3-azetidinyl]amino]-2-oxoethylidene]amino]oxy]-2-methyl-propanoicacid), metronidazole(2-methyl-5-nitro-1H-imidazole-1-ethanol), epiroprim(5-[[3,5-diethoxy-4-(1H-pyrrol-1-yl)phenyl]methyl]-2,4-pyrimidinediamine),OCA-983(1-[[(2S)-2-amino-3-methyl-1-oxobutyl]amino]-2,5-anhydro-3-S-[(4R,5S,6S)-2-carboxy-6-[(1R)-1-hydxoxyethyl]-4-methyl-7-oxo-1-azabicyclo[3.2.0]hept-2-en-3-yl]-1,4-dideoxy-3-thio-D-threo-pentitol),GV-143253 (trinem), sanfetrinem ((1S, 5S, 8aS, 8bR)-1,2, 5, 6, 7, 8, 8a,8b-octahydro-1-[(1R)-1-hydroxyethyl]-5-methoxy-2-oxo-azeto[2,1-a]isoindole-4-carboxylicacid), CS-834 ((4R, 5S,6S)-6-[(IR)-1-hydroxyethyl]-4-methyl-7-oxo-3-[[((3R)-5-oxo-3-pyrrolidinyl]thio]-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylicacid (2,2-dimethyl-1-oxopropoxy)methyl ester), biapenem(6-[[(4R,5S,6S)-2-carboxy-6-[(1R)-1-hydroxyethyl]-4-methyl-7-oxo-1-azabicyclo[3.2.0]hept-2-en-3-yl]thio]-6,7-dihydro-5H-pyrazolo[1,2-a][1,2,4]triazol-4-iuminner salt), KA 159 (stipiamide), dynemicin A((1S,4R,4aR,14S,14aS,18Z)-1,4,7,12,13,14-hexahydro-6,8,11-trihydroxy-3-methoxy-1-methyl-7,12-dioxo-4a,14a-epoxy-4,14-[3]hexene[1,5]diynonaphtho[2,3-c]phenanthridine-2-carboxylicacid), DX8739((4R,5S,6S)-3-[[(3S,5S)-5-[[4-[(2S)-5-amino-2-hydroxy-1-oxopentyl]-1-piperazinyl]carbonyl]-3-pyrrolidinyl]thio]-6-[(1R)-1-hydroxyethyl]-4-methyl-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylicacid), DU 6681((4R,5S,6S)-3-[[(6S)-6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-6-yl]thio]-6-[(1R)-1-hydroxyethyl]-4-methyl-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylicacid), cefluprenam((2E)-N-(2-amino-2-oxoethyl)-3-[(6R,7R)-7-[[(2Z)-(5-amino-1,2,4-thiadiazol-3-yl)[(fluoromethoxy)imino]acetyl]amino]-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl]-N-ethyl-N-methyl-2-propen-1-aminiuminner salt), ER 35786((4R,5S,6S)-6-[(1R)-1-hydroxyethyl]-3-[[(3S,5S)-5-[(R)-hydroxy(3R)-3-pyrrolidinylmethyl]-3-pyrrolidinyl]thio]-4-methyl-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylicacid monohydrochloride), cefoselis((6R,7R)-7-[[(2Z)-(2-amino-4-thiazolyl)(methoxyimino)acetyl]amino]-3-[[2,3-dihydro-2-(2-hydroxyethyl)-3-imino-1H-pyrazol-1-yl]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid), sanfetrinem celexetil((1S,5S,8aS,8bR)-1,2,5,6,7,8,8a,8b-octabydro-1-[(1R)-1-hydroxyethyl]-5-methoxy-2-oxo-azeto[2,1-a]isoindole-4-carboxylicacid 1-[(cyclohexyloxy)carbonyl]oxy]ethyl ester), cefpirome(1-[[(6R,7R)-7-[[(2Z)-(2-amino-4-thiazolyl)(methoxyimino)acetyl]amino]-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl]methyl]-6,7-dihydro-5H-cyclopenta[b]pyridiniuminner salt), HMR-3647(3-de[(2,6-dideoxy-3-C-methyl-3-O-methyl-alpha-L-ribo-hexopyranosyl)oxy]-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl[[4-(4-(3-pyridinyl)-1H-imidazol-1-yl]butyl]imino]]-erythromycin),RU-59863 (C-7 catechol substituted cephalosporin), KP 736((6R,7R)-7-[[(2Z)-(2-amino-4-thiazolyl)[[(1,4-dihydro-1,5-dihydroxy-4-oxo-2-pyridinyl)methoxy]imino]acetyl]amino]-8-oxo-3-[(1,2,3-thiadiazol-5-ylthio)methyl]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid disodium salt), Rifalazil(1′,4-didehydro-1-deoxy-1,4-dihydro-3′-hydroxy-5′-[4-(2-methylpropyl)-1-piperazinyl]-1-oxo-rifamycinVIII), MEN 10700((5R,6S)-3-[[(2-amino-2-oxoethyl)methylamino]methyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-4-thia-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylicacid), lenapenem((4R,5S,6S)-6-[(1R)-1-hydroxyethyl]-3-[[(3S,5S)-5-[(1R)-1-hydroxy-3-(methylamino)propyl]-3-pyrrolidinyl]thio]-4-methyl-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylicacid), BO 2502A((4R,5S,6S)-3-[(2S,3′S,4S)-[2,3′-bipyrrolidin]-4-ylthio]-6-[(1R)-1-hydroxyethyl]-4-methyl-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylicacid), NE-1530 (3′-sialyllacto-N-neotetraose), PR 39(L-arginyl-L-arginyl-L-arginyl-L-prolyl-L-arginyl-L-prolyl-L-prolyl-L-tryosyl-L-leucyl-L-prolyl-L-arginyl-L-prolyl-L-arginyl-L-prolyl-L-prolyl-L-prolyl-L-phenylalanyl-L-phenylalanyl-L-prolyl-L-prolyl-L-arginyl-L-leucyl-L-prolyl-L-prolyl-L-arginyl-L-isoleucyl-L-prolyl-L-prolylglycyl-L-phenylalanyl-L-prolyl-L-prolyl-L-arginyl-L-phenylalanyl-L-prolyl-L-prolyl-L-arginyl-L-phenylalanyl-L-prolinamide[SEQ ID NO: 1[), K130(5-[[4-[3-[[4-[(4-aminophenyl)sulfonyl]phenyl]amino]propoxy]-3,5-dimethoxyphenyl]methyl]-2,4-pyrimidinediamine),PD 138312((R)-7-[3-(1-amino-1-methylethyl)-1-pyrrolidinyl]-1-cyclopropyl-6-fluoro-1,4-dihydro-4-oxo-1,8-naphthyridine-3-carboxylicacid), PD 140248(7-[(3R)-3-[(1S)-1-aminoethyl]-1-pyrrolidinyl]-1-(2,4-difluorophenyl)-6-fluoro-1,4-dihydro-4-oxo-1,8-naphthyridine-3-carboxylicacid), CP 111905(5-deoxy-5-[[(2E)-3-[3-hydroxy-4-(2-propenyloxy)phenyl]-2-methyl-1-oxo-2-propenyl]amino]-1,2-O-mehyle-D-neo-inositol),sulopenem((5R,6S)-6-[(1R)-1-hydroxyethyl]-7-oxo-3-[[(1R,3S)-tetrahydro-1-oxido-3-thienyl]thio]-4-thia-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylicacid), ritipenam acoxyl((5R,6R)-3-[[(aminocarbonyl)oxy]methyl]-6-[(1R)-1-hydroxyethyl]-7-oxo-4-thia-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylicacid (acetyloxy)methyl ester), RO-65-5788((6R,7R)-7-[[(2Z)-(5-amino-1,2,4-thiadiazol-3-yl)(hydroxyimino)acetyl]amino]-3-[(E)-[(3′R)-1′-[[(5-methyl-2-oxo-1,3-dioxol-4-yl)methoxy]carbonyl]-2-oxo[1,3′-bipyrrolidin]-3-ylidene]methyl]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid monosodium salt), Sch-40832(N-[[48-[1-[[2,6-dideoxy-3-O-(2,6-dideoxy-D-arabino-hexopyranosyl)-D-arabino-hexopyranosyl]oxy]ethyl]-15-ethylidene-1,3a,4,5,10,11,12,13,14,15,19,20,21,22,28,29,41,42-octadecahydro-41-hydroxy-12,45-bis(1-hydroxyethyl)-1-(hydroxymethyl)-22-(1-hydroxy-1-methylpropyl)-36-methyl-51,54,57-tris(methylene)-3-(methylthio)-10,13,20,27,38,49,52,55,58-nanaoxo-18H,27H-5a,29-(iminoethaniminoethaniminoethaniminoethanimino[7,2]quinolinomethanoxymethano)-9,6:19,16:26,23:33,30-tetranitrilo-16H,33aH-imidazo[1′,5′:1,6]pyrido[3,2-m][1,11,17,24,4,7,20,27]tetrathiatetraazacyclotriacontin-1-yl]carbonyl]-2,3-didehydroalanyl-2,3-didehydro-alaninemethyl ester stereoisomer), micacocidin A((OC-6-26-A)-[(4S)-2-[(2S)-2-[(2R,4R)-2-[(4R)-4,5-dihydro-2-[2-(hydroxy-.kappa.O)-6-pentylphenyl]-4-thiazolyl-.kappa.N3]-3-methyl-4-thiazolidinyl-.kappa.N3]-2-(hydroxy-.kappa.O)-1,1-dimethylethyl]-4,5-dihydro-4-methyl-4-thiazolecarboxylato(2-)-.kappa.N3,.kappa.O4]-Zinc), SR-15402((1S,5S,8aS,8bR)-1,2,5,6,7,8,8a,8b-octahydro-1-[(1R)-1-hydroxyethyl]-2-oxo-5-[(3S)-3-pyrrolidinylthio]-azeto[2,1-a]isoindole-4-carboxylicacid TOC 39(1-(2-amino-2-oxoethyl)-4-[[(1E)-2-[(6R,7R)-7-[[(2Z)-(2-amino-4-thiazolyl)(hydroxyimino)acetyl]amino]-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl]ethenyl]thio]-pyridiniuminner salt), carumonam([[(Z)-[2-[[(2S,3S)-2-[[(aminocarbonyl)oxy]methyl]-4-oxo-1-sulfo-3-azetidinyl]amino]-1-(2-amino-4-thiazolyl)-2-oxoethylidene]amino]oxy]-aceticacid), cefozopran(1-[[(6R,7R)-7-[[(2Z)-5-amino-1,2,4-thiadiazol-3-yl)(methoxyimino)acetyl]amino]-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl]methyl]-imidazo[1,2-b]pyridaziniuminner salt), cefetamet pivoxil((6R,7R)-7-[[(2Z)-(2-amino-4-thiazolyl)(methoxyimino)acetyl]amino]-3-methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylicacid (2,2-dimethyl-1-oxopropoxy)methyl ester), and T 3811(des-F(6)-quinolone).
 20. The method according to claim 18, wherein saidantimicrobial agent is selected from the group consisting of imipenen,amikacin, netilmicin, fosfomycin, gentamicin, celhiaxone, teicoplanin,ZIRACIN(56-deacetyl-57-demethyl-45-O-de(2-methyl-1-oxopropyl)-12-O-(2,3,6-trideoxy-3-C-methyl-4-O-methyl-3-nitro-alpha-L-arabino-hexopyranosyl)flambamycin),LY333328 (oritavancin), HMR-3647(3-de[(2,6-dideoxy-3-C-methyl-3-O-methyl-alpha-L-ribo-hexopyranosyl)oxy]-11,12-dideoxy-6-O-methyl-3-oxo-12,11-[oxycarbonyl[[4-[4-(3-pyridinyl)-1H-imidazol-1-yl-]butyl]imino]]-erythromycin),linezolid(N-[[(5S)-3-[3fluoro-4-(4-morpholinyl)phenyl]-2-oxo-5-oxazolidinyl]methyl]acetamide),SYNERCID (dalfopristin-quinupristin), aztreonam(2-[[(Z)-[1-(2-amino-4-thiazolyl)-2-[[(2S,3S)-2-methyl-4-oxo-1sulfo-3-azetidinyl]amino]-2-oxoethylidene]amino]oxy-2-methyl-propanoicacid), and metronidazole (2-methyl-5-nitro-1H-imidazole-1-ethanol). 21.The method according to claim 13, wherein said subject is selected fromthe group consisting of a human and an animal.
 22. The method accordingto claim 21, wherein said subject is a human.
 23. The compound of claim1 having the formula (II):

wherein R⁵⁶ is an optionally substituted straight-chain C₈-C₁₄ alkylgroup.
 24. A method of using the compound according to claim 23 to makea compound according to either of claim 1 or 2 of the formula:

wherein said method comprises treating a compound of claim 23 with areagent selected from the group consisting of an isocyanates,isothiocyanates, activatcd esters, acid chlorides, sulfonylchlorides,activated sulfonamides, heterocycles bearing readily displaceablegroups, imidates, and lactones; or alternatively, treating a compound ofclaim 23 reductively with an aldehyde.
 25. The compound according toeither of claim 1 or 2 wherein said compound is selected from Cpd # R R¹R² 1 NHCONH(CH₂)₇CH₃ NH₂

2 NHCONH(CH₂)₁₁CH₃ NH₂

3 NHCONH(CH₂)₁₀CH₃

5

17 NHCONH(CH₂)₁₁CH₃

48 NHCONH(CH₂)₁₀CH₃ NH₂

56 NHCONH(CH₂)₇CH₃

57 NHCONH(CH₂)₁₀CH₃

58 NHCONH(CH₂)₁₁CH₃

62 NHCONH(CH₂)₇CH₃

63 NHCONH(CH₂)₁₀CH₃

64 NHCONH(CH₂)₁₁CH₃

69 NHCONH(CH₂)₇CH₃

70 NHCONH(CH₂)₇CH₃

71 NHCONH(CH₂)₇CH₃

75 NHCONH(CH₂)₁₀CH₃

76 NHCONH(CH₂)₇CH₃

77 NHCONH(CH₂)₇CH₃

78 NHCONH(CH₂)₇CH₃

87 NHCONH(CH₂)₁₁CH₃

88 NHCONH(CH₂)₁₁CH₃

89 NHCONH(CH₂)₁₁CH₃

108 NHCONH(CH₂)₁₀CH₃

113 NHCONH(CH₂)₁₀CH₃

114 NHCONH(CH₂)₁₀CH₃

117 NHCONH(CH₂)₈CH₃ NHBoc

118 NHCONH(CH₂)₈CH₃ NH₂

119 NHCONH(CH₂)₉CH₃ NHBoc

120 NHCONH(CH₂)₉CH₃ NH₂


26. The compound according to claim 25 wherein said compound is selectedfrom Cpd # R R¹ R² 2 NHCONH(CH₂)₁₁CH₃ NH₂

3 NHCONH(CH₂)₁₀CH₃

48 NHCONH(CH₂)₁₀CH₃ NH₂

89 NHCONH(CH₂)₁₁CH₃

118 NHCONH(CH₂)₈CH₃ NH₂

120 NHCONH(CH₂)₉CH₃ NH₂


27. The compound according claim 2, wherein R is selected from the groupconsisting of:

wherein each of R³ and R⁵ is independently selected from the groupconsisting of hydrido, alkyl, aryl, heterocyclyl and heteroaryl, andwherein R²⁰⁰ is aryl.
 28. The compound accordiug to claim 27, wherein Ris

and wherein R^(4′) is a substituted phenyl.
 29. The compound accordingto claim 28, wherein R is

and wherein X³ is chloro or trifluoromethyl.
 30. The method according toclaim 19, wherein anti-folate agents are sulfonamides.
 31. The methodaccording to claim 18, wherein the second antimicrobial agent is asynthetic antibacterial selected from the group consisting ofnitrofurans, methenamine mandelate and methenamine hippurate.